LOCUS       pRHA-113.        5837 bp DNA     circular SYN 01-JAN-1980
DEFINITION  pBR322-derived low copy number plasmid encoding the rhaB promoter 
            plus rhaR and rhaS genes, for rhamnose-inducible protein expression 
            in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    pRHA-113
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5837)
  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 5837)
  AUTHORS   Xbrane Bioscience
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5837)
  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..5837
                     /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..2127
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             3391..3579
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3684..3824
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4010..4598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4772..5629)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5630..5734)
                     /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 gggtacctcg accgatgccc ttgagagcct tcaacccagt
     2161 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     2221 tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg
     2281 ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc
     2341 cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat
     2401 tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg
     2461 ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt
     2521 gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggatcgct
     2581 cgcggctctt accagcctaa cttcgatcat tggaccgctg atcgtcacgg cgatttatgc
     2641 cgcctcggcg agcacatgga acgggttggc atggattgta ggcgccgccc tataccttgt
     2701 ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc
     2761 cggcggcacc tcgctaacgg attcaccact ccaagaattg gagccaatca attcttgcgg
     2821 agaactgtga atgcgcaaac caacccttgg cagaacatat ccatcgcgtc cgccatctcc
     2881 agcagccgca cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc
     2941 gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat
     3001 gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga
     3061 gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca
     3121 gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga
     3181 acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc
     3241 cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca
     3301 tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg
     3361 aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac
     3421 atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac
     3481 gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc
     3541 agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     3601 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     3661 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
     3721 tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     3781 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct
     3841 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3901 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3961 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4021 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4081 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4141 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4201 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4261 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4321 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4381 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4441 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4501 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4561 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4621 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4681 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4741 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4801 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4861 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4921 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4981 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5041 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt
     5101 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5161 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5221 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5281 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5341 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg
     5401 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5461 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5521 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5581 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5641 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5701 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5761 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     5821 cctttcgtct tcaagaa
//
