LOCUS       pRHA-109.        4193 bp DNA     circular SYN 01-JAN-1980
DEFINITION  pBR322-derived high copy number plasmid encoding the rhaB promoter 
            plus rhaR and rhaS genes, for rhamnose-inducible protein expression 
            in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    pRHA-109
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4193)
  AUTHORS   Giacalone MJ, Gentile AM, Lovitt BT, Berkley NL, Gunderson CW, 
            Surber MW.
  TITLE     Toxic protein expression in Escherichia coli using a rhamnose-based 
            tightly regulated and tunable promoter system.
  JOURNAL   BioTechniques 2006;40:355-64.
  PUBMED    16568824
REFERENCE   2  (bases 1 to 4193)
  AUTHORS   Xbrane Bioscience
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4193)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "2006"; volume: "40"; pages: "355-64"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     This plasmid sequence was reconstructed from the procedure described
            in the original paper. The plasmid size is slightly different than 
            reported.
FEATURES             Location/Qualifiers
     source          1..4193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(40..885)
                     /label=rhaR
                     /note="transcriptional activator of rhaR and rhaS"
     CDS             complement(962..1795)
                     /label=rhaS
                     /note="positive regulator of the rhaB promoter"
     promoter        1941..2059
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     misc_feature    2077..2132
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     rep_origin      complement(2366..2954)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3128..3985)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3986..4090)
                     /label=AmpR promoter
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gcttaattaa tctttctgcg aattgagatg
       61 acgccactgg ctgggcgtca tcccggtttc ccgggtaaac accaccgaaa aatagttact
      121 atcttcaaag ccacattcgg tcgaaatatc actgattaac aggcggctat gctggagaag
      181 atattgcgca tgacacactc tgacctgtcg cagatattga ttgatggtca ttccagtctg
      241 ctggcgaaat tgctgacgca aaacgcgctc actgcacgat gcctcatcac aaaatttatc
      301 cagcgcaaag ggacttttca ggctagccgc cagccgggta atcagcttat ccagcaacgt
      361 ttcgctggat gttggcggca acgaatcact ggtgtaacga tggcgattca gcaacatcac
      421 caactgcccg aacagcaact cagccatttc gttagcaaac ggcacatgct gactactttc
      481 atgctcaagc tgaccgataa cctgccgcgc ctgcgccatc cccatgctac ctaagcgcca
      541 gtgtggttgc cctgcgctgg cgttaaatcc cggaatcgcc ccctgccagt caagattcag
      601 cttcagacgc tccgggcaat aaataatatt ctgcaaaacc agatcgttaa cggaagcgta
      661 ggagtgttta tcgtcagcat gaatgtaaaa gagatcgcca cgggtaatgc gataagggcg
      721 atcgttgagt acatgcaggc cattaccgcg ccagacaatc accagctcac aaaaatcatg
      781 tgtatgttca gcaaagacat cttgcggata acggtcagcc acagcgactg cctgctggtc
      841 gctggcaaaa aaatcatctt tgagaagttt taactgatgc gccaccgtgg ctacctcggc
      901 cagagaacga agttgattat tcgcaatatg gcgtacaaat acgttgagaa gattcgcgtt
      961 attgcagaaa gccatcccgt ccctggcgaa tatcacgcgg tgaccagtta aactctcggc
     1021 gaaaaagcgt cgaaaagtgg ttactgtcgc tgaatccaca gcgataggcg atgtcagtaa
     1081 cgctggcctc gctgtggcgt agcagatgtc gggctttcat cagtcgcagg cggttcaggt
     1141 atcgctgagg cgtcagtccc gtttgctgct taagctgccg atgtagcgta cgcagtgaaa
     1201 gagaaaattg atccgccacg gcatcccaat tcacctcatc ggcaaaatgg tcctccagcc
     1261 aggccagaag caagttgaga cgtgatgcgc tgttttccag gttctcctgc aaactgcttt
     1321 tacgcagcaa gagcagtaat tgcataaaca agatctcgcg actggcggtc gagggtaaat
     1381 cattttcccc ttcctgctgt tccatctgtg caaccagctg tcgcacctgc tgcaatacgc
     1441 tgtggttaac gcgccagtga gacggatact gcccatccag ctcttgtggc agcaactgat
     1501 tcagcccggc gagaaactga aatcgatccg gcgagcgata cagcacattg gtcagacaca
     1561 gattatcggt atgttcatac agatgccgat catgatcgcg tacgaaacag accgtgccac
     1621 cggtgatggt atagggctgc ccattaaaca catgaatacc cgtgccatgt tcgacaatca
     1681 caatttcatg aaaatcatga tgatgttcag gaaaatccgc ctgcgggagc cggggttcta
     1741 tcgccacgga cgcgttacca gacggaaaaa aatccacact atgtaatacg gtcatactgg
     1801 cctcctgatg tcgtcaacac ggcgaaatag taatcacgag gtcaggttct taccttaaat
     1861 tttcgacgga aaaccacgta aaaaacgtcg atttttcaag atacagcgtg aattttcagg
     1921 aaatgcggtg agcatcacat caccacaatt cagcaaattg tgaacatcat cacgttcatc
     1981 tttccctggt tgccaatggc ccattttcct gtcagtaacg agaaggtcgc gaattcaggc
     2041 gctttttaga ctggtcgtaa tgaaattcag gaggttgtcg actctagagg atccccgcgc
     2101 cctcatccga aagggcgtat gggtaccata tgcggtgtga aataccgcac agatgcgtaa
     2161 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2221 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2281 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2341 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2401 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2461 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2521 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2581 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2641 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2701 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2761 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     2821 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2881 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2941 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3001 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3061 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3121 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3181 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3241 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3301 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3361 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3421 gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3481 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3541 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3601 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3661 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3721 gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag
     3781 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3841 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3901 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     3961 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4021 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4081 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     4141 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaa
//
