LOCUS       40924_10806        5618 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Promoter vector pCK302, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5618)
  AUTHORS   Kelly CL, Liu Z, Yoshihara A, Jenkinson SF, Wormald MR, Otero JM, 
            Estevez AM, Kato A, Marqvorsen MH, Fleet GW, Estevez RJ, Izumori K, 
            Heap JT.
  TITLE     Synthetic chemical inducers and genetic decoupling enable orthogonal
            control of the rhaBAD promoter
  JOURNAL   ACS Synth Biol (2016) In press
  PUBMED    27247275
REFERENCE   2  (bases 1 to 5618)
  AUTHORS   Kelly CL, Liu Z, Yoshihara A, Jenkinson SF, Wormald MR, Otero J, 
            Estevez A, Kato A, Marqvorsen MHS., Fleet GWJ., Estevez RJ, Izumori 
            K, Heap JT.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-JAN-2016) Centre for Synthetic Biology and Innovation,
            Department of Life Sciences, Imperial College London, South 
            Kensington Campus, London SW7 2AZ, United Kingdom
REFERENCE   3  (bases 1 to 5618)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5618)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-JAN-2016) Centre for Synthetic Biology and Innovation, 
            Department of Life Sciences, Imperial College London, South 
            Kensington Campus, London SW7 2AZ, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     875..891
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      908..1021
                     /label=rpn txn term terminator
                     /note="rpn txn term terminator"
                     /regulatory_class="terminator"
     regulatory      1028..1328
                     /label=bla txn term terminator
                     /note="bla txn term terminator"
                     /regulatory_class="terminator"
     promoter        1345..1462
                     /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)"
     CDS             1518..2231
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     CDS             2250..2267
                     /label=6xHis
                     /note="6xHis affinity tag"
     regulatory      2300..2474
                     /label=rrnB1 B2 T1 term terminator
                     /note="rrnB1 B2 T1 term terminator"
                     /regulatory_class="terminator"
     promoter        2561..2652
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      2583..2611
                     /label=Pamp promoter
                     /note="Pamp promoter"
                     /regulatory_class="promoter"
     CDS             2653..3510
                     /label=AmpR
                     /note="beta-lactamase"
     regulatory      3525..3558
                     /note="Translation initiation rate of 20000 using Salis RBS
                     calculator; MG1655 rhaS RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             3559..4392
                     /label=rhaS
                     /note="positive regulator of the rhaB promoter"
     CDS             4467..5546
                     /label=lacI
                     /note="lac repressor"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttagccca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      661 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      721 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      781 cggaaggcga gagtagggaa ctgccaggca tcaaactaag cagaaggccc ctgacggatg
      841 gcctttttgc gtttctacaa actctttctg tgttgtaaaa cgacggccag tcttaagctc
      901 gggccccctg ggcggttctg ataacgagta atcgttaatc cgcaaataac gtaaaaaccc
      961 gcttcggcgg gtttttttat ggggggagtt tagggaaaga gcatttgtca gaatatttaa
     1021 gggcgcctgt cactttgctt gatatatgag aattatttaa ccttataaat gagaaaaaag
     1081 caacgcactt taaataagat acgttgcttt ttcgattgat gaacacctat aattaaacta
     1141 ttcatctatt atttatgatt ttttgtatat acaatatttc tagtttgtta aagagaatta
     1201 agaaaataaa tctcgaaaat aataaaggga aaatcagttt ttgatatcaa aattatacat
     1261 gtcaacgata atacaaaata taatacaaac tataagatgt tatcagtatt tattatgcat
     1321 ttagaataaa ttttgtgtcg cccttccaca attcagcaaa ttgtgaacat catcacgttc
     1381 atctttccct ggttgccaat ggcccatttt cctgtcagta acgagaaggt cgcgtattca
     1441 ggcgcttttt agactggtcg taatgaaacg gtttccctct agaaataatt ttgtttaact
     1501 tttaggaggt aaaacatatg cgtaaaggcg aagagctgtt cactggtgtc gtccctattc
     1561 tggtggaact ggatggtgat gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag
     1621 gtgacgcaac taatggtaaa ctgacgctga agttcatctg tactactggt aaactgccgg
     1681 taccttggcc gactctggta acgacgctga cttatggtgt tcagtgcttt gctcgttatc
     1741 cggaccatat gaagcagcat gacttcttca agtccgccat gccggaaggc tatgtgcagg
     1801 aacgcacgat ttcctttaag gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg
     1861 aaggcgatac cctggtaaac cgcattgagc tgaaaggcat tgactttaaa gaagacggca
     1921 atatcctggg ccataagctg gaatacaatt ttaacagcca caatgtttac atcaccgccg
     1981 ataaacaaaa aaatggcatt aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca
     2041 gcgtgcagct ggctgatcac taccagcaaa acactccaat cggtgatggt cctgttctgc
     2101 tgccagacaa tcactatctg agcacgcaaa gcgttctgtc taaagatccg aacgagaaac
     2161 gcgatcatat ggttctgctg gagttcgtaa ccgcagcggg catcacgcat ggtatggatg
     2221 aactgtacaa aggctccggc tccggctccc accatcacca tcaccattga taactcgagc
     2281 cccaagggcg acaccccata attagcccgg gcgaaaggcc cagtctttcg actgagcctt
     2341 tcgttttatt tgatgcctgg cagttcccta ctctcgcatg gggagtcccc acactaccat
     2401 cggcgctacg gcgtttcact tctgagttcg gcatggggtc aggtgggacc accgcgctac
     2461 tgccgccagg caaacaaggg gtgttatgag ccatattcag gtataaatgg gctcgcgata
     2521 atgttcagaa ttggttaatt ggttgtaaca ctgaccccta tttgtttatt tttctaaata
     2581 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2641 aaaaggaaga atatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2701 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2761 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2821 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2881 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2941 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3001 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3061 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3121 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3181 gacaccacga tgcctgtagc gatggcaaca acgttgcgca aactattaac tggcgaacta
     3241 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3301 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc cggagccggt
     3361 gagcgtggtt ctcgcggtat catcgcagcg ctggggccag atggtaagcc ctcccgtatc
     3421 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3481 gagataggtg cctcactgat taagcattgg taagcggcgc gccatatttc gccgtgttga
     3541 cgacatcagg aggccagtat gaccgtatta catagtgtgg atttttttcc gtctggtaac
     3601 gcgtccgtgg cgatagaacc ccggctcccg caggcggatt ttcctgaaca tcatcatgat
     3661 tttcatgaaa ttgtgattgt cgaacatggc acgggtattc atgtgtttaa tgggcagccc
     3721 tataccatca ccggtggcac ggtctgtttc gtacgcgatc atgatcggca tctgtatgaa
     3781 cataccgata atctgtgtct gaccaatgtg ctgtatcgct cgccggatcg atttcagttt
     3841 ctcgccgggc tgaatcagtt gctgccacaa gagctggatg ggcagtatcc gtctcactgg
     3901 cgcgttaacc acagcgtatt gcagcaggtg cgacagctgg ttgcacagat ggaacagcag
     3961 gaaggggaaa atgatttacc ctcgaccgcc agtcgcgaga tcttgtttat gcaattactg
     4021 ctcttgctgc gtaaaagcag tttgcaggag aacctggaaa acagcgcatc acgtctcaac
     4081 ttgcttctgg cctggctgga ggaccatttt gccgatgagg tgaattggga tgccgtggcg
     4141 gatcaatttt ctctttcact gcgtacgcta catcggcagc ttaagcagca aacgggactg
     4201 acgcctcagc gatacctgaa ccgcctgcga ctgatgaaag cccgacatct gctacgccac
     4261 agcgaggcca gcgttactga catcgcctat cgctgtggat tcagcgacag taaccacttt
     4321 tcgacgcttt ttcgccgaga gtttaactgg tcaccgcgtg atattcgcca gggacgggat
     4381 ggctttctgc aataatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg
     4441 aagagagtca attcagggtg gtgaatatga aaccagtaac gttatacgat gtcgcagagt
     4501 atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg
     4561 cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg
     4621 tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg
     4681 ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg
     4741 ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc
     4801 acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg
     4861 atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg
     4921 accagacacc catcaacagt attattttct cccatgagga cggtacgcga ctgggcgtgg
     4981 agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg
     5041 tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc
     5101 cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa
     5161 tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg
     5221 gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg
     5281 gatacgacga taccgaagat agctcatgtt atatcccgcc gttaaccacc atcaaacagg
     5341 attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg
     5401 cggtgaaggg caatcagctg ttgccagtct cactggtgaa aagaaaaacc accctggcgc
     5461 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     5521 aggtttcccg actggaaagc gggcagtgac tcatgaccaa aatcccttaa cgtgagttac
     5581 gcgcgcgtcg ttccactgag cgtcagaccc cgtagaaa
//
