LOCUS       40924_10841        5930 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle expression vector pCK324, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5930)
  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 5930)
  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 5930)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5930)
  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..5930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     1..1015
                     /label=similar to ndhB
                     /note="similar to ndhB"
     regulatory      1026..1059
                     /label=synthetic
                     /note="synthetic"
                     /regulatory_class="ribosome_binding_site"
     CDS             1060..1776
                     /label=EYFP
                     /note="enhanced YFP"
     regulatory      1786..1895
                     /label=kan
                     /note="kan"
                     /regulatory_class="promoter"
     CDS             1896..2708
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      2712..2734
                     /label=synthetic
                     /note="synthetic"
                     /regulatory_class="ribosome_binding_site"
     CDS             2735..3568
                     /label=rhaS
                     /note="positive regulator of the rhaB promoter"
     misc_recomb     3572..4610
                     /label=similar to ndhB
                     /note="similar to ndhB"
     rep_origin      4943..5488
                     /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 gtgaatcggg taagtttata atatacaaag gaggtagaaa tggtgagcaa gggcgaggag
     1081 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1141 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1201 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
     1261 ggcctgcaat gcttcgcccg ctaccccgac cacatgaagc tgcacgactt cttcaagtcc
     1321 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1381 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1441 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1501 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1561 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1621 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     1681 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1741 gccgggatca ctctcggcat ggacgagctg tacaagtaag gatccaaagc cacgttgtgt
     1801 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     1861 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     1921 cttgctcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     1981 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg
     2041 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     2101 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     2161 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg
     2221 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     2281 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     2341 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     2401 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct
     2461 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     2521 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     2581 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     2641 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     2701 agtttttcta agagcctttc gaagcagcaa tcaaatgacc gtattacata gtgtggattt
     2761 ttttccgtct ggtaacgcgt ccgtggcgat agaaccccgg ctcccgcagg cggattttcc
     2821 tgaacatcat catgattttc atgaaattgt gattgtcgaa catggcacgg gtattcatgt
     2881 gtttaatggg cagccctata ccatcaccgg tggcacggtc tgtttcgtac gcgatcatga
     2941 tcggcatctg tatgaacata ccgataatct gtgtctgacc aatgtgctgt atcgctcgcc
     3001 ggatcgattt cagtttctcg ccgggctgaa tcagttgctg ccacaagagc tggatgggca
     3061 gtatccgtct cactggcgcg ttaaccacag cgtattgcag caggtgcgac agctggttgc
     3121 acagatggaa cagcaggaag gggaaaatga tttaccctcg accgccagtc gcgagatctt
     3181 gtttatgcaa ttactgctct tgctgcgtaa aagcagtttg caggagaacc tggaaaacag
     3241 cgcatcacgt ctcaacttgc ttctggcctg gctggaggac cattttgccg atgaggtgaa
     3301 ttgggatgcc gtggcggatc aattttctct ttcactgcgt acgctacatc ggcagcttaa
     3361 gcagcaaacg ggactgacgc ctcagcgata cctgaaccgc ctgcgactga tgaaagcccg
     3421 acatctgcta cgccacagcg aggccagcgt tactgacatc gcctatcgct gtggattcag
     3481 cgacagtaac cacttttcga cgctttttcg ccgagagttt aactggtcac cgcgtgatat
     3541 tcgccaggga cgggatggct ttctgcaata atcttggggg aaattaagac caaactcgat
     3601 gacctccaaa aagatgtaac ttctcttaag atcgatatgg caacggtgaa aaccgagtta
     3661 tctgcggtca ggatggagat aggtacagtc aaggatgatg ttaaagatgt caaagggcgg
     3721 gctaatgctc aaatttgggc gttgattctt gccgtcatcg gagccataat taccaccttg
     3781 gtgcgttttg gcattttccc taatccctaa caaaaaagcg accaggcttt tctttcaatt
     3841 gcccgatcgc ctttgatatt ttcccaaagg ataaaagcta gtccattcag aatcgagcct
     3901 taaagtactc ccatattggc tagccccaga attactccag cgccgaggat gtggccaaag
     3961 ctagcggtgc ccagcacagc ccctaaacca aagccgccaa agaagttaga ggaaggcatg
     4021 ggggtgccca cattttgttg tttgatggtc aatttaccaa aggcgatcgc caaaatgttg
     4081 caagcaatca tcaccccagc aactttaggg ctccaggaca gggtggcggg aacggcggtg
     4141 gccaacaaaa agctatgcat tgagattctc cagaataaag acggttttta aagggatagc
     4201 cccacgctaa tgggggtctt taaaaatctc atcttacggg gacgctctgc ccctgggaaa
     4261 ccaccgttgc aatacttaac aaattttcgt ttttagcttg gcaaatgtct ttggcaaaat
     4321 tggttgatct ggcttaaatc gtcagttatt tgccctggaa tagtctgggg acgggcaatt
     4381 ctgatcagat ttacccccaa cgcttccgcc actttttgct taaccaattc tcccccctgg
     4441 gcaccggagg ctttagttac caccccttga atttgccatt gttgccacag ggctttttcc
     4501 aatggttcgg ctacgggggg gcgcaaagca atgatacggt cggaagtaaa cccagcggcg
     4561 atcgcctggg ctagggcttg gggatagggc agaatacggg caaatagaac aaatctggag
     4621 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4681 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4741 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4801 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4861 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4921 accccttaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg
     4981 aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt
     5041 tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc
     5101 cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc
     5161 agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg
     5221 cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac
     5281 ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc
     5341 ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg gaaacgcctg
     5401 gtatctttat agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg
     5461 cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc gcggccctct cacttccctg
     5521 ttaagtatct tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc catttccgct
     5581 cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga atatatcctg
     5641 tatcacatat tctgctgacg caccggtgca gccttttttc tcctgccaca tgaagcactt
     5701 cactgacacc ctcatcagtg ccaacatagt aagccagtat acactccgct agcgctgagg
     5761 tctgcctcgt gaagaaggtg ttgctgactc ataccaggcc tgaatcgccc catcatccag
     5821 ccagaaagtg agggagccac ggttgatgag agctttgttg taggtggacc agttggtgat
     5881 tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg ggaagatgcg
//
