LOCUS       40924_10811        5188 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle expression vector pCK305, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5188)
  AUTHORS   Kelly CL, Taylor GM, Hitchcock A, Torres-Mendez A, Heap JT.
  TITLE     A Rhamnose-Inducible System for Precise and Temporal Control of Gene
            Expression in Cyanobacteria
  JOURNAL   ACS Synth Biol (2018) In press
  PUBMED    29544054
REFERENCE   2  (bases 1 to 5188)
  AUTHORS   Kelly CL, Taylor GM, Mendez AT, Hitchcock A, Heap JT.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-OCT-2017) 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 5188)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5188)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-OCT-2017) 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..5188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     1..1015
                     /label=similar to ndhB
                     /note="similar to ndhB"
     promoter        1021..1138
                     /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)"
     regulatory      1144..1177
                     /label=synthetic
                     /note="synthetic"
                     /regulatory_class="ribosome_binding_site"
     CDS             1178..1894
                     /label=EYFP
                     /note="enhanced YFP"
     regulatory      1904..2013
                     /label=kan
                     /note="kan"
                     /regulatory_class="promoter"
     CDS             2014..2826
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     misc_recomb     2830..3868
                     /label=similar to ndhB
                     /note="similar to ndhB"
     rep_origin      4201..4746
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
ORIGIN
        1 gtgggtcaat ccctcggttt ggcgatcgcc ttggtgtttg tcattgctgg tattgccttt
       61 aaaatttccg ctgtaccctt ccaccaatgg acccccgacg tttatgaagg ttctcccacc
      121 ccggtggtgg cttttctttc ggtgggctct aaggcggccg gttttgccgt agctatccga
      181 ttactagtga ctgcctttgg tggtattacc gatgagtggc acgttatttt caccgctctg
      241 gccgtgttga gcatggtgct gggcaacgtg gtggctttag cccaaaccag catgaaacgg
      301 atgttggcct actcttccat cggtcaagca ggctttgtga tgattggcct agtggccggc
      361 agtgaagatg gttacgccag catggttttc tacatgctca tctatctgtt tatgaacctg
      421 ggggcgttta gttgcattat tctcttcacc ctccgcactg gcagtgacca aattagtgat
      481 tacgctggtc tgtaccacaa agaccccttg ttaaccttgg gcttgagcat ttgtttatta
      541 tccttggggg gcattcctcc tctggcgggc tttttcggca aaatttacat cttctgggcc
      601 ggttggcaat cgggattgta tggcctagtc ctacttggtc tggttaccag tgtagtttcc
      661 atctactact acatccgggt ggtgaaaatg atggtggtga aggagcccca ggaaatgtcc
      721 gaagtaatca aaaattaccc ggccatcaaa tggaatttac ccggcatgcg tcccctacag
      781 gtgggcattg tcgctacttt ggttgctacc tcgctggcag gtattctggc taatcccctc
      841 tttaacctcg ccaccgattc cgtggtcagc accaagatgt tgcagacagc cctccagcaa
      901 acaggagaaa ctccggcgat cgccatttcc catgatttac cctaggggta tcaggaaata
      961 ttgctttgca ggcaaaagcc aatgagtgta actatagaaa ccgatttaaa ggagactgca
     1021 gccacaattc agcaaattgt gaacatcatc acgttcatct ttccctggtt gccaatggcc
     1081 cattttcctg tcagtaacga gaaggtcgcg tattcaggcg ctttttagac tggtcgtaat
     1141 gaatcgggta agtttataat atacaaagga ggtagaaatg gtgagcaagg gcgaggagct
     1201 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1261 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1321 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg
     1381 cctgcaatgc ttcgcccgct accccgacca catgaagctg cacgacttct tcaagtccgc
     1441 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1501 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1561 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1621 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1681 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1741 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct
     1801 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1861 cgggatcact ctcggcatgg acgagctgta caagtaagga tccaaagcca cgttgtgtct
     1921 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     1981 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct
     2041 tgctcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     2101 cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg
     2161 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     2221 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     2281 actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc attccaggta
     2341 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     2401 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     2461 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     2521 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataagctttt gccattctca
     2581 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     2641 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     2701 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     2761 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     2821 tttttctaat cttgggggaa attaagacca aactcgatga cctccaaaaa gatgtaactt
     2881 ctcttaagat cgatatggca acggtgaaaa ccgagttatc tgcggtcagg atggagatag
     2941 gtacagtcaa ggatgatgtt aaagatgtca aagggcgggc taatgctcaa atttgggcgt
     3001 tgattcttgc cgtcatcgga gccataatta ccaccttggt gcgttttggc attttcccta
     3061 atccctaaca aaaaagcgac caggcttttc tttcaattgc ccgatcgcct ttgatatttt
     3121 cccaaaggat aaaagctagt ccattcagaa tcgagcctta aagtactccc atattggcta
     3181 gccccagaat tactccagcg ccgaggatgt ggccaaagct agcggtgccc agcacagccc
     3241 ctaaaccaaa gccgccaaag aagttagagg aaggcatggg ggtgcccaca ttttgttgtt
     3301 tgatggtcaa tttaccaaag gcgatcgcca aaatgttgca agcaatcatc accccagcaa
     3361 ctttagggct ccaggacagg gtggcgggaa cggcggtggc caacaaaaag ctatgcattg
     3421 agattctcca gaataaagac ggtttttaaa gggatagccc cacgctaatg ggggtcttta
     3481 aaaatctcat cttacgggga cgctctgccc ctgggaaacc accgttgcaa tacttaacaa
     3541 attttcgttt ttagcttggc aaatgtcttt ggcaaaattg gttgatctgg cttaaatcgt
     3601 cagttatttg ccctggaata gtctggggac gggcaattct gatcagattt acccccaacg
     3661 cttccgccac tttttgctta accaattctc ccccctgggc accggaggct ttagttacca
     3721 ccccttgaat ttgccattgt tgccacaggg ctttttccaa tggttcggct acgggggggc
     3781 gcaaagcaat gatacggtcg gaagtaaacc cagcggcgat cgcctgggct agggcttggg
     3841 gatagggcag aatacgggca aatagaacaa atctggagcc ggtgagcgtg ggtctcgcgg
     3901 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3961 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4021 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4081 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4141 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccttaataa gatgatcttc
     4201 ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca
     4261 gggcggtttt tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg
     4321 aggagcgcag tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa
     4381 gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt
     4441 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg
     4501 gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg
     4561 gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca
     4621 ggagagcgca cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg
     4681 tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta
     4741 tggaaaaacg gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt
     4801 ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc
     4861 gtagcgagtc agtgagcgag gaagcggaat atatcctgta tcacatattc tgctgacgca
     4921 ccggtgcagc cttttttctc ctgccacatg aagcacttca ctgacaccct catcagtgcc
     4981 aacatagtaa gccagtatac actccgctag cgctgaggtc tgcctcgtga agaaggtgtt
     5041 gctgactcat accaggcctg aatcgcccca tcatccagcc agaaagtgag ggagccacgg
     5101 ttgatgagag ctttgttgta ggtggaccag ttggtgattt tgaacttttg ctttgccacg
     5161 gaacggtctg cgttgtcggg aagatgcg
//
