LOCUS       40924_10836        6048 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle expression vector pCK321, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6048)
  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 6048)
  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 6048)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6048)
  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..6048
                     /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"
     regulatory      2830..2852
                     /label=synthetic
                     /note="synthetic"
                     /regulatory_class="ribosome_binding_site"
     CDS             2853..3686
                     /label=rhaS
                     /note="positive regulator of the rhaB promoter"
     misc_recomb     3690..4728
                     /label=similar to ndhB
                     /note="similar to ndhB"
     rep_origin      5061..5606
                     /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 tttttctaag agcctttcga agcagcaatc aaatgaccgt attacatagt gtggattttt
     2881 ttccgtctgg taacgcgtcc gtggcgatag aaccccggct cccgcaggcg gattttcctg
     2941 aacatcatca tgattttcat gaaattgtga ttgtcgaaca tggcacgggt attcatgtgt
     3001 ttaatgggca gccctatacc atcaccggtg gcacggtctg tttcgtacgc gatcatgatc
     3061 ggcatctgta tgaacatacc gataatctgt gtctgaccaa tgtgctgtat cgctcgccgg
     3121 atcgatttca gtttctcgcc gggctgaatc agttgctgcc acaagagctg gatgggcagt
     3181 atccgtctca ctggcgcgtt aaccacagcg tattgcagca ggtgcgacag ctggttgcac
     3241 agatggaaca gcaggaaggg gaaaatgatt taccctcgac cgccagtcgc gagatcttgt
     3301 ttatgcaatt actgctcttg ctgcgtaaaa gcagtttgca ggagaacctg gaaaacagcg
     3361 catcacgtct caacttgctt ctggcctggc tggaggacca ttttgccgat gaggtgaatt
     3421 gggatgccgt ggcggatcaa ttttctcttt cactgcgtac gctacatcgg cagcttaagc
     3481 agcaaacggg actgacgcct cagcgatacc tgaaccgcct gcgactgatg aaagcccgac
     3541 atctgctacg ccacagcgag gccagcgtta ctgacatcgc ctatcgctgt ggattcagcg
     3601 acagtaacca cttttcgacg ctttttcgcc gagagtttaa ctggtcaccg cgtgatattc
     3661 gccagggacg ggatggcttt ctgcaataat cttgggggaa attaagacca aactcgatga
     3721 cctccaaaaa gatgtaactt ctcttaagat cgatatggca acggtgaaaa ccgagttatc
     3781 tgcggtcagg atggagatag gtacagtcaa ggatgatgtt aaagatgtca aagggcgggc
     3841 taatgctcaa atttgggcgt tgattcttgc cgtcatcgga gccataatta ccaccttggt
     3901 gcgttttggc attttcccta atccctaaca aaaaagcgac caggcttttc tttcaattgc
     3961 ccgatcgcct ttgatatttt cccaaaggat aaaagctagt ccattcagaa tcgagcctta
     4021 aagtactccc atattggcta gccccagaat tactccagcg ccgaggatgt ggccaaagct
     4081 agcggtgccc agcacagccc ctaaaccaaa gccgccaaag aagttagagg aaggcatggg
     4141 ggtgcccaca ttttgttgtt tgatggtcaa tttaccaaag gcgatcgcca aaatgttgca
     4201 agcaatcatc accccagcaa ctttagggct ccaggacagg gtggcgggaa cggcggtggc
     4261 caacaaaaag ctatgcattg agattctcca gaataaagac ggtttttaaa gggatagccc
     4321 cacgctaatg ggggtcttta aaaatctcat cttacgggga cgctctgccc ctgggaaacc
     4381 accgttgcaa tacttaacaa attttcgttt ttagcttggc aaatgtcttt ggcaaaattg
     4441 gttgatctgg cttaaatcgt cagttatttg ccctggaata gtctggggac gggcaattct
     4501 gatcagattt acccccaacg cttccgccac tttttgctta accaattctc ccccctgggc
     4561 accggaggct ttagttacca ccccttgaat ttgccattgt tgccacaggg ctttttccaa
     4621 tggttcggct acgggggggc gcaaagcaat gatacggtcg gaagtaaacc cagcggcgat
     4681 cgcctgggct agggcttggg gatagggcag aatacgggca aatagaacaa atctggagcc
     4741 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4801 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4861 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4921 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4981 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5041 cccttaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa
     5101 aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac caactctttg
     5161 aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct
     5221 taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag
     5281 tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5341 gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc
     5401 ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg
     5461 taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt
     5521 atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct
     5581 tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt
     5641 aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg
     5701 ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta
     5761 tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca
     5821 ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag cgctgaggtc
     5881 tgcctcgtga agaaggtgtt gctgactcat accaggcctg aatcgcccca tcatccagcc
     5941 agaaagtgag ggagccacgg ttgatgagag ctttgttgta ggtggaccag ttggtgattt
     6001 tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcg
//
