LOCUS       40924_10831        6058 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle expression vector pCK320, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6058)
  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 6058)
  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 6058)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6058)
  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..6058
                     /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..2862
                     /label=synthetic
                     /note="synthetic"
                     /regulatory_class="ribosome_binding_site"
     CDS             2863..3696
                     /label=rhaS
                     /note="positive regulator of the rhaB promoter"
     misc_recomb     3700..4738
                     /label=similar to ndhB
                     /note="similar to ndhB"
     rep_origin      5071..5616
                     /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 gcccctatat ttcaccttcg agattacccg ttatgaccgt attacatagt
     2881 gtggattttt ttccgtctgg taacgcgtcc gtggcgatag aaccccggct cccgcaggcg
     2941 gattttcctg aacatcatca tgattttcat gaaattgtga ttgtcgaaca tggcacgggt
     3001 attcatgtgt ttaatgggca gccctatacc atcaccggtg gcacggtctg tttcgtacgc
     3061 gatcatgatc ggcatctgta tgaacatacc gataatctgt gtctgaccaa tgtgctgtat
     3121 cgctcgccgg atcgatttca gtttctcgcc gggctgaatc agttgctgcc acaagagctg
     3181 gatgggcagt atccgtctca ctggcgcgtt aaccacagcg tattgcagca ggtgcgacag
     3241 ctggttgcac agatggaaca gcaggaaggg gaaaatgatt taccctcgac cgccagtcgc
     3301 gagatcttgt ttatgcaatt actgctcttg ctgcgtaaaa gcagtttgca ggagaacctg
     3361 gaaaacagcg catcacgtct caacttgctt ctggcctggc tggaggacca ttttgccgat
     3421 gaggtgaatt gggatgccgt ggcggatcaa ttttctcttt cactgcgtac gctacatcgg
     3481 cagcttaagc agcaaacggg actgacgcct cagcgatacc tgaaccgcct gcgactgatg
     3541 aaagcccgac atctgctacg ccacagcgag gccagcgtta ctgacatcgc ctatcgctgt
     3601 ggattcagcg acagtaacca cttttcgacg ctttttcgcc gagagtttaa ctggtcaccg
     3661 cgtgatattc gccagggacg ggatggcttt ctgcaataat cttgggggaa attaagacca
     3721 aactcgatga cctccaaaaa gatgtaactt ctcttaagat cgatatggca acggtgaaaa
     3781 ccgagttatc tgcggtcagg atggagatag gtacagtcaa ggatgatgtt aaagatgtca
     3841 aagggcgggc taatgctcaa atttgggcgt tgattcttgc cgtcatcgga gccataatta
     3901 ccaccttggt gcgttttggc attttcccta atccctaaca aaaaagcgac caggcttttc
     3961 tttcaattgc ccgatcgcct ttgatatttt cccaaaggat aaaagctagt ccattcagaa
     4021 tcgagcctta aagtactccc atattggcta gccccagaat tactccagcg ccgaggatgt
     4081 ggccaaagct agcggtgccc agcacagccc ctaaaccaaa gccgccaaag aagttagagg
     4141 aaggcatggg ggtgcccaca ttttgttgtt tgatggtcaa tttaccaaag gcgatcgcca
     4201 aaatgttgca agcaatcatc accccagcaa ctttagggct ccaggacagg gtggcgggaa
     4261 cggcggtggc caacaaaaag ctatgcattg agattctcca gaataaagac ggtttttaaa
     4321 gggatagccc cacgctaatg ggggtcttta aaaatctcat cttacgggga cgctctgccc
     4381 ctgggaaacc accgttgcaa tacttaacaa attttcgttt ttagcttggc aaatgtcttt
     4441 ggcaaaattg gttgatctgg cttaaatcgt cagttatttg ccctggaata gtctggggac
     4501 gggcaattct gatcagattt acccccaacg cttccgccac tttttgctta accaattctc
     4561 ccccctgggc accggaggct ttagttacca ccccttgaat ttgccattgt tgccacaggg
     4621 ctttttccaa tggttcggct acgggggggc gcaaagcaat gatacggtcg gaagtaaacc
     4681 cagcggcgat cgcctgggct agggcttggg gatagggcag aatacgggca aatagaacaa
     4741 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4801 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4861 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4921 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4981 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5041 agcgtcagac cccttaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc
     5101 ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac
     5161 caactctttg aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc
     5221 agtttagcct taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg
     5281 ctgctgccag tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg
     5341 ataaggcgca gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa
     5401 ctgcctaccc ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga
     5461 atgacaccgg taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga
     5521 aacgcctggt atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt
     5581 tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca
     5641 cttccctgtt aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca
     5701 tttccgctcg ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat
     5761 atatcctgta tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg
     5821 aagcacttca ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag
     5881 cgctgaggtc tgcctcgtga agaaggtgtt gctgactcat accaggcctg aatcgcccca
     5941 tcatccagcc agaaagtgag ggagccacgg ttgatgagag ctttgttgta ggtggaccag
     6001 ttggtgattt tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcg
//
