LOCUS       pRHA-67.        4745 bp DNA     circular SYN 01-JAN-1980
DEFINITION  pUC18-derived plasmid encoding the rhaB promoter plus rhaR and rhaS 
            genes, for rhamnose-inducible protein expression in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    pRHA-67
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4745)
  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 4745)
  AUTHORS   Xbrane Bioscience
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4745)
  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..4745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     gene            405..410
                     /gene="rhaT"
                     /gene_synonym="ECK3900; JW3878"
                     /locus_tag="b3907"
                     /label=rhaT
                     /note="rhaT"
                     /db_xref="EcoGene:EG11313"
                     /db_xref="GeneID:948398"
     CDS             complement(410..1255)
                     /label=rhaR
                     /note="transcriptional activator of rhaR and rhaS"
     CDS             complement(1332..2165)
                     /label=rhaS
                     /note="positive regulator of the rhaB promoter"
     promoter        2311..2429
                     /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    2439..2444
                     /label=RBS
                     /note="RBS"
                     /note="ribosome binding site"
     misc_feature    2447..2508
                     /label=MCS
                     /note="MCS"
                     /note="pUC19 multiple cloning site"
     primer_bind     complement(2524..2540)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2548..2564)
                     /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)."
     promoter        complement(2572..2602)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2617..2638)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2926..3514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3688..4545)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4546..4650)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttaattaa tctttctgcg
      421 aattgagatg acgccactgg ctgggcgtca tcccggtttc ccgggtaaac accaccgaaa
      481 aatagttact atcttcaaag ccacattcgg tcgaaatatc actgattaac aggcggctat
      541 gctggagaag atattgcgca tgacacactc tgacctgtcg cagatattga ttgatggtca
      601 ttccagtctg ctggcgaaat tgctgacgca aaacgcgctc actgcacgat gcctcatcac
      661 aaaatttatc cagcgcaaag ggacttttca ggctagccgc cagccgggta atcagcttat
      721 ccagcaacgt ttcgctggat gttggcggca acgaatcact ggtgtaacga tggcgattca
      781 gcaacatcac caactgcccg aacagcaact cagccatttc gttagcaaac ggcacatgct
      841 gactactttc atgctcaagc tgaccgataa cctgccgcgc ctgcgccatc cccatgctac
      901 ctaagcgcca gtgtggttgc cctgcgctgg cgttaaatcc cggaatcgcc ccctgccagt
      961 caagattcag cttcagacgc tccgggcaat aaataatatt ctgcaaaacc agatcgttaa
     1021 cggaagcgta ggagtgttta tcgtcagcat gaatgtaaaa gagatcgcca cgggtaatgc
     1081 gataagggcg atcgttgagt acatgcaggc cattaccgcg ccagacaatc accagctcac
     1141 aaaaatcatg tgtatgttca gcaaagacat cttgcggata acggtcagcc acagcgactg
     1201 cctgctggtc gctggcaaaa aaatcatctt tgagaagttt taactgatgc gccaccgtgg
     1261 ctacctcggc cagagaacga agttgattat tcgcaatatg gcgtacaaat acgttgagaa
     1321 gattcgcgtt attgcagaaa gccatcccgt ccctggcgaa tatcacgcgg tgaccagtta
     1381 aactctcggc gaaaaagcgt cgaaaagtgg ttactgtcgc tgaatccaca gcgataggcg
     1441 atgtcagtaa cgctggcctc gctgtggcgt agcagatgtc gggctttcat cagtcgcagg
     1501 cggttcaggt atcgctgagg cgtcagtccc gtttgctgct taagctgccg atgtagcgta
     1561 cgcagtgaaa gagaaaattg atccgccacg gcatcccaat tcacctcatc ggcaaaatgg
     1621 tcctccagcc aggccagaag caagttgaga cgtgatgcgc tgttttccag gttctcctgc
     1681 aaactgcttt tacgcagcaa gagcagtaat tgcataaaca agatctcgcg actggcggtc
     1741 gagggtaaat cattttcccc ttcctgctgt tccatctgtg caaccagctg tcgcacctgc
     1801 tgcaatacgc tgtggttaac gcgccagtga gacggatact gcccatccag ctcttgtggc
     1861 agcaactgat tcagcccggc gagaaactga aatcgatccg gcgagcgata cagcacattg
     1921 gtcagacaca gattatcggt atgttcatac agatgccgat catgatcgcg tacgaaacag
     1981 accgtgccac cggtgatggt atagggctgc ccattaaaca catgaatacc cgtgccatgt
     2041 tcgacaatca caatttcatg aaaatcatga tgatgttcag gaaaatccgc ctgcgggagc
     2101 cggggttcta tcgccacgga cgcgttacca gacggaaaaa aatccacact atgtaatacg
     2161 gtcatactgg cctcctgatg tcgtcaacac ggcgaaatag taatcacgag gtcaggttct
     2221 taccttaaat tttcgacgga aaaccacgta aaaaacgtcg atttttcaag atacagcgtg
     2281 aattttcagg aaatgcggtg agcatcacat caccacaatt cagcaaattg tgaacatcat
     2341 cacgttcatc tttccctggt tgccaatggc ccattttcct gtcagtaacg agaaggtcgc
     2401 gaattcaggc gctttttaga ctggtcgtaa tgaaattcag gaggttgtcg actctagagg
     2461 atccccgcgc cctcatccga aagggcgtat gggtacggta ccgagctcga attcgtaatc
     2521 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2581 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2641 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2701 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2761 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2821 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2881 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2941 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3001 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3061 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3121 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3181 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3241 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3301 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3361 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3421 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3481 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3541 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3601 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3661 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3721 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3781 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3841 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3901 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3961 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4021 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4081 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4141 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4201 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4261 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4321 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4381 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4441 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4501 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4561 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4621 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     4681 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     4741 tcgtc
//
