LOCUS       40924_10821        5156 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle expression vector pCK313, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5156)
  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 5156)
  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 5156)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5156)
  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..5156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     1..1015
                     /label=similar to ndhB
                     /note="similar to ndhB"
     regulatory      1022..1111
                     /note="rhamnose; minus CRP-binding site"
                     /regulatory_class="promoter"
     misc_binding    1025..1041
                     /label=RhaS binding site
                     /bound_moiety="RhaS"
                     /note="operator 1"
     misc_binding    1058..1073
                     /label=RhaS binding site
                     /bound_moiety="RhaS"
                     /note="operator 2"
     regulatory      1071..1076
                     /label=PrhaBAD
                     /note="PrhaBAD"
                     /regulatory_class="minus_35_signal"
     regulatory      1095..1100
                     /label=PrhaBAD
                     /note="PrhaBAD"
                     /regulatory_class="minus_10_signal"
     regulatory      1112..1145
                     /label=synthetic
                     /note="synthetic"
                     /regulatory_class="ribosome_binding_site"
     CDS             1146..1862
                     /label=EYFP
                     /note="enhanced YFP"
     regulatory      1872..1981
                     /label=kan
                     /note="kan"
                     /regulatory_class="promoter"
     CDS             1982..2794
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     misc_recomb     2798..3836
                     /label=similar to ndhB
                     /note="similar to ndhB"
     rep_origin      4169..4714
                     /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 gttcatcttt ccctggttgc caatggccca ttttcctgtc agtaacgaga aggtcgcgta
     1081 ttcaggcgct ttttagactg gtcgtaatga atcgggtaag tttataatat acaaaggagg
     1141 tagaaatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1201 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1261 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1321 ccaccctcgt gaccaccttc ggctacggcc tgcaatgctt cgcccgctac cccgaccaca
     1381 tgaagctgca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1441 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1501 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1561 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1621 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1681 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1741 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1801 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1861 agtaaggatc caaagccacg ttgtgtctca aaatctctga tgttacattg cacaagataa
     1921 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     1981 tatgagccat attcaacggg aaacgtcttg ctcgaggccg cgattaaatt ccaacatgga
     2041 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2101 ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2161 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2221 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2281 gatccccggg aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2341 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2401 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2461 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2521 agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2581 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2641 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2701 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     2761 attgcagttt catttgatgc tcgatgagtt tttctaatct tgggggaaat taagaccaaa
     2821 ctcgatgacc tccaaaaaga tgtaacttct cttaagatcg atatggcaac ggtgaaaacc
     2881 gagttatctg cggtcaggat ggagataggt acagtcaagg atgatgttaa agatgtcaaa
     2941 gggcgggcta atgctcaaat ttgggcgttg attcttgccg tcatcggagc cataattacc
     3001 accttggtgc gttttggcat tttccctaat ccctaacaaa aaagcgacca ggcttttctt
     3061 tcaattgccc gatcgccttt gatattttcc caaaggataa aagctagtcc attcagaatc
     3121 gagccttaaa gtactcccat attggctagc cccagaatta ctccagcgcc gaggatgtgg
     3181 ccaaagctag cggtgcccag cacagcccct aaaccaaagc cgccaaagaa gttagaggaa
     3241 ggcatggggg tgcccacatt ttgttgtttg atggtcaatt taccaaaggc gatcgccaaa
     3301 atgttgcaag caatcatcac cccagcaact ttagggctcc aggacagggt ggcgggaacg
     3361 gcggtggcca acaaaaagct atgcattgag attctccaga ataaagacgg tttttaaagg
     3421 gatagcccca cgctaatggg ggtctttaaa aatctcatct tacggggacg ctctgcccct
     3481 gggaaaccac cgttgcaata cttaacaaat tttcgttttt agcttggcaa atgtctttgg
     3541 caaaattggt tgatctggct taaatcgtca gttatttgcc ctggaatagt ctggggacgg
     3601 gcaattctga tcagatttac ccccaacgct tccgccactt tttgcttaac caattctccc
     3661 ccctgggcac cggaggcttt agttaccacc ccttgaattt gccattgttg ccacagggct
     3721 ttttccaatg gttcggctac gggggggcgc aaagcaatga tacggtcgga agtaaaccca
     3781 gcggcgatcg cctgggctag ggcttgggga tagggcagaa tacgggcaaa tagaacaaat
     3841 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3901 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3961 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4021 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4081 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4141 cgtcagaccc cttaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt
     4201 gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca
     4261 actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag
     4321 tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct
     4381 gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat
     4441 aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact
     4501 gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata acagcggaat
     4561 gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc cagggggaaa
     4621 cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc
     4681 gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact
     4741 tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt
     4801 tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaatat
     4861 atcctgtatc acatattctg ctgacgcacc ggtgcagcct tttttctcct gccacatgaa
     4921 gcacttcact gacaccctca tcagtgccaa catagtaagc cagtatacac tccgctagcg
     4981 ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac caggcctgaa tcgccccatc
     5041 atccagccag aaagtgaggg agccacggtt gatgagagct ttgttgtagg tggaccagtt
     5101 ggtgattttg aacttttgct ttgccacgga acggtctgcg ttgtcgggaa gatgcg
//
