LOCUS       40924_30835        6032 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range expression vector pMHE5Tc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6032)
  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 6032)
  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 6032)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6032)
  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..6032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        187..205
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    206..230
                     /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             245..267
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    274..276
                     /label=fusion protein start codon
                     /note="fusion protein start codon"
     CDS             280..297
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             298..309
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     CDS             364..387
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             391..414
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     terminator      482..529
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1163..1822)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(1823..2592)
                     /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    2816..3809
                     /note="RP4 mob; oriT"
     regulatory      complement(3899..4014)
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"
     CDS             complement(4323..5510)
                     /label=TcR
                     /note="tetracycline efflux protein"
     regulatory      5895..6010
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"
ORIGIN
        1 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
       61 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
      121 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
      181 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
      241 taattttgtt taactttaag aaggagatat accatggggc atcatcatca tcatcatatc
      301 gagggaaggc ctgtcgacgg tatcgataag cttgatatcg aattcctgca gctcgaggga
      361 tccgactaca aggacgacga cgacaagaac tggagccatc cgcagttcga gaagtgaaga
      421 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
      481 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
      541 aactatatcc ggatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggaaattgt
      601 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
      661 ccaataggcc gactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg
      721 gtgcccttaa acgcctggtg ctacgcctga ataagtgata ataagcggat gaatggcaga
      781 aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt
      841 atgtctattg ctggtttacc ggtttattga ctaccggaag cagtgtgacc gtgtgcttct
      901 caaatgcctg aggccagttt gctcaggctc tccccgtgga ggtaataatt gacgatatga
      961 tcatttattc tgcctcccag agcctgataa aaacggtgaa tccgttagcg aggtgccgcc
     1021 ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca
     1081 gacaaggtat agggcggcga ggcggctaca gccgatagtc tggaacagcg cacttacggg
     1141 ttgctgcgca acccaagtgc taccggcgcg gcagcgtgac ccgtgtcggc ggctccaacg
     1201 gctcgccatc gtccagaaaa cacggctcat cgggcatcgg caggcgctgc tgcccgcgcc
     1261 gttcccattc ctccgtttcg gtcaaggctg gcaggtctgg ttccatgccc ggaatgccgg
     1321 gctggctggg cggctcctcg ccggggccgg tcggtagttg ctgctcgccc ggatacaggg
     1381 tcgggatgcg gcgcaggtcg ccatgcccca acagcgattc gtcctggtcg tcgtgatcaa
     1441 ccaccacggc ggcactgaac accgacaggc gcaactggtc gcggggctgg ccccacgcca
     1501 cgcggtcatt gaccacgtag gccgacacgg tgccggggcc gttgagcttc acgacggaga
     1561 tccagcgctc ggccaccaag tccttgactg cgtattggac cgtccgcaaa gaacgtccga
     1621 tgagcttgga aagtgtcttc tggctgacca ccacggcgtt ctggtggccc atctgcgcca
     1681 cgaggtgatg cagcagcatt gccgccgtgg gtttcctcgc aataagcccg gcccacgcct
     1741 catgcgcttt gcgttccgtt tgcacccagt gaccgggctt gttcttggct tgaatgccga
     1801 tttctctgga ctgcgtggcc atgcttatct ccatgcggta gggtgccgca cggttgcggc
     1861 accatgcgca atcagctgca acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa
     1921 gtggcagtca attacagatt ttctttaacc tacgcaatga gctattgcgg ggggtgccgc
     1981 aatgagctgt tgcgtacccc ccttttttaa gttgttgatt tttaagtctt tcgcatttcg
     2041 ccctatatct agttctttgg tgcccaaaga agggcacccc tgcggggttc ccccacgcct
     2101 tcggcgcggc tccccctccg gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc
     2161 ggccttcggc cttgcccaag gtggcgctgc ccccttggaa cccccgcact cgccgccgtg
     2221 aggctcgggg ggcaggcggg cgggcttcgc cttcgactgc ccccactcgc ataggcttgg
     2281 gtcgttccag gcgcgtcaag gccaagccgc tgcgcggtcg ctgcgcgagc cttgacccgc
     2341 cttccacttg gtgtccaacc ggcaagcgaa gcgcgcaggc cgcaggccgg aggcttttcc
     2401 ccagagaaaa ttaaaaaaat tgatggggca aggccgcagg ccgcgcagtt ggagccggtg
     2461 ggtatgtggt cgaaggctgg gtagccggtg ggcaatccct gtggtcaagc tcgtgggcag
     2521 gcgcagcctg tccatcagct tgtccagcag ggttgtccac gggccgagcg aagcgagcca
     2581 gccggtggcc gctcgcggcc atcgtccaca tatccacggg ctggcaaggg agcgcagcga
     2641 ccgcgcaggg cgaagcccgg agagcaagcc cgtagggcgc cgcagccgcc gtaggcggtc
     2701 acgactttgc gaagcaaagt ctagtgagta tactcaagca ttgagtggcc cgccggaggc
     2761 accgccttgc gctgcccccg tcgagccggt tggacaccaa aagggagggg caggcatggc
     2821 ggcatacgcg atcatgcgat gcaagaagct ggcgaaaatg ggcaacgtgg cggccagtct
     2881 caagcacgcc taccgcgagc gcgagacgcc caacgctgac gccagcagga cgccagagaa
     2941 cgagcactgg gcggccagca gcaccgatga agcgatgggc cgactgcgcg agttgctgcc
     3001 agagaagcgg cgcaaggacg ctgtgttggc ggtcgagtac gtcatgacgg ccagcccgga
     3061 atggtggaag tcggccagcc aagaacagca ggcggcgttc ttcgagaagg cgcacaagtg
     3121 gctggcggac aagtacgggg cggatcgcat cgtgacggcc agcatccacc gtgacgaaac
     3181 cagcccgcac atgaccgcgt tcgtggtgcc gctgacgcag gacggcaggc tgtcggccaa
     3241 ggagttcatc ggcaacaaag cgcagatgac ccgcgaccag accacgtttg cggccgctgt
     3301 ggccgatcta gggctgcaac ggggcatcga gggcagcaag gcacgtcaca cgcgcattca
     3361 ggcgttctac gaggccctgg agcggccacc agtgggccac gtcaccatca gcccgcaagc
     3421 ggtcgagcca cgcgcctatg caccgcaggg attggccgaa aagctgggaa tctcaaagcg
     3481 cgttgagacg ccggaagccg tggccgaccg gctgacaaaa gcggttcggc aggggtatga
     3541 gcctgcccta caggccgccg caggagcgcg tgagatgcgc aagaaggccg atcaagccca
     3601 agagacggcc cgagaccttc gggagcgcct gaagcccgtt ctggacgccc tggggccgtt
     3661 gaatcgggat atgcaggcca aggccgccgc gatcatcaag gccgtgggcg aaaagctgct
     3721 gacggaacag cgggaagtcc agcgccagaa acaggcccag cgccagcagg aacgcgggcg
     3781 cgcacatttc cccgaaaagt gccacctggc ggcgttgtga caatttaccg aacaactccg
     3841 cggccgggaa gccgatctcg gcttgaacga attgttaggt ggcggtactt gggtcgatag
     3901 ctttatgctt gtaaaccgtt ttgtgaaaaa atttttaaaa taaaaaaggg gacctctagg
     3961 gtccccaatt aattagtaat ataatctatt aaaggtcatt caaaaggtca tccaccggat
     4021 caattcccct gctcgcgcag gctgggtgcc aagctctcgg gtaacatcaa ggcccgatcc
     4081 ttggagccct tgccctcccg cacgatgatc gtgccgtgat cgaaatccag atccttgacc
     4141 cgcagttgca aaccctcact gatccgccga gatgcgccgc gtgcggctgc tggagatggc
     4201 ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa
     4261 ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt
     4321 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat
     4381 agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc
     4441 gccgtgacga tcagcggtcc agtgatcgaa gttaggctgg taagagccgc gagcgatcct
     4501 tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc
     4561 atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc
     4621 gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc
     4681 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     4741 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     4801 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     4861 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     4921 aaggctctca agggcatcgg tcgacgctct cccttatgcg actcctgcat taggaagcag
     4981 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     5041 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     5101 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     5161 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     5221 cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag tagcgaagcg
     5281 agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg tgcgcataga
     5341 aattgcatca acgcatatag cgctagcagc acgccatagt gactggcgat gctgtcggaa
     5401 tggacgatat cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca
     5461 tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc
     5521 tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagctcgcg cagatcagtt
     5581 ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataatgtct
     5641 aacaattcgt tcaagccgac gccgcttcgc ggcgcggctt aactcaagcg ttagatgcac
     5701 taagcacata attgctcaca gccaaactat caggtcaagt ctgcttttat tatttttaag
     5761 cgtgcataat aagccctaca caaattggga gatatatcat gaaaggctgg ctttttcttg
     5821 ttatcgcaat agttggcgaa gtaatcgcaa catccgcatt aaaatctagc gagggcttta
     5881 ctaagctgat ccggtggatg accttttgaa tgacctttaa tagattatat tactaattaa
     5941 ttggggaccc tagaggtccc cttttttatt ttaaaaattt tttcacaaaa cggtttacaa
     6001 gcataaagct caaggagatg gcgcccaaca gt
//
