LOCUS       40924_27114        6973 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Transposon delivery vector pKUT-Tn5-Cm DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6973)
  AUTHORS   Watabe K, Mimuro M, Tsuchiya T.
  TITLE     Development of a highly-frequent in vivo transposon mutagenesis 
            system for Synechocystis sp. PCC 6803 and Synechococcus sp. PCC 7942
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6973)
  AUTHORS   Tsuchiya T, Watabe K.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-JUN-2014) Contact:Tohru Tsuchiya Kyoto University; 
            Yoshida-nihonmatsu-cho, Sakyo-ku, Kyoto, Kyoto 606-8501, Japan
REFERENCE   3  (bases 1 to 6973)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6973)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-JUN-2014) Contact:Tohru Tsuchiya Kyoto University; 
            Yoshida-nihonmatsu-cho, Sakyo-ku, Kyoto, Kyoto 606-8501, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..19
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     CDS             complement(118..774)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(775..877)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     misc_feature    complement(1000..1018)
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     protein_bind    1030..1046
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     rep_origin      complement(1081..1469)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(2090..2458)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(2491..2600)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(3681..4538)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4539..4643)
                     /label=AmpR promoter
     terminator      complement(4697..4733)
                     /label=soxR terminator
                     /note="bidirectional E. coli soxR transcription terminator"
     terminator      complement(4755..4782)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(4874..4960)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(5171..6598)
                     /codon_start=1
                     /label=Tn5 transposase
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAAQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTLPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     protein_bind    complement(6616..6632)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6640..6668)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
ORIGIN
        1 ctgtctctta tacacatcta cgcgtgtccg aatttctgcc attcatccgc ttattatcac
       61 ttattcaggc gtagcaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc
      121 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
      181 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
      241 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
      301 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
      361 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
      421 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
      481 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
      541 tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
      601 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
      661 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
      721 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
      781 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
      841 attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt
      901 ggcccagggc ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc
      961 ttccgtcaca ggtatttatt cggacacgcg tgcggccgca gatgtgtata agagacagaa
     1021 ttccacaaat tgttatccgc tcacaattcc acatgtggaa ttccacatgt ggaattccca
     1081 tgtcagccgt taagtgttcc tgtgtcactc aaaattgctt tgagaggctc taagggcttc
     1141 tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta aaccttaaaa gctttaaaag
     1201 ccttatatat tctttttttt cttataaaac ttaaaacctt agaggctatt taagttgctg
     1261 atttatatta attttattgt tcaaacatga gagcttagta cgtgaaacat gagagcttag
     1321 tacgttagcc atgagagctt agtacgttag ccatgagggt ttagttcgtt aaacatgaga
     1381 gcttagtacg ttaaacatga gagcttagta cgtgaaacat gagagcttag tacgtactat
     1441 caacaggttg aactgctgat cttcagatcc tctacgccgg acgcatcgtg gccggatcca
     1501 gccgaccagg ctttccacgc ccgcgtgccg ctccatgtcg ttcgcgcggt tctcggaaac
     1561 gcgctgccgc gtttcgtgat tgtcacgctc aagcccgtag tcccgttcga gcgtcgcgca
     1621 gaggtcagcg agggcgcggt aggcccgata cggctcatgg atggtgtttc gggtcgggtg
     1681 aatcttgttg atggcgatat ggatgtgcag gttgtcggtg tcgtgatgca cggcactgac
     1741 gcgctgatgc tcggcgaagc caagcccagc gcagatgcgg tcctcaatcg cgcgcaacgt
     1801 ctccgcgtcg ggcttctctc ccgcgcggaa gctaaccagc aggtgatagg tcttgtcggc
     1861 ctcggaacgg gtgttgccgt gctgggtcgc catcacctcg gccatgacag cgggcagggt
     1921 gtttgcctcg cagttcgtga cgcgcacgtg acccaggcgc tcggtcttgc cttgctcgtc
     1981 ggtgatgtac ttcaccagct ccgcgaagtc gctcttcttg atggagcgca tggggacgtg
     2041 cttggcaatc acgcgcaccc cccggccgtt ttagcggcta aaaaagtcat ggctctgccc
     2101 tcgggcggac cacgcccatc atgaccttgc caagctcgtc ctgcttctct tcgatcttcg
     2161 ccagcagggc gaggatcgtg gcatcaccga accgcgccgt gcgcgggtcg tcggtgagcc
     2221 agagtttcag caggccgccc aggcggccca ggtcgccatt gatgcgggcc agctcgcgga
     2281 cgtgctcata gtccacgacg cccgtgattt tgtagccctg gccgacggcc agcaggtagg
     2341 ccgacaggct catgccggcc gccgccgcct tttcctcaat cgctcttcgt tcgtctggaa
     2401 ggcagtacac cttgataggt gggctgccct tcctggttgg cttggtttca tcagccatcc
     2461 gcttgccctc atctgttacg ccggcggtag ccggccagcc tcgcagagca ggattcccgt
     2521 tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc gcgggtgggc
     2581 ctacttcacc tatcctgccc ggctgacgcc gttggataca ccaaggaaag tctacacgaa
     2641 ccctttggca aaatcctgta tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta
     2701 taatgacccc gaagcagggt tatgcagcgg aaaagcgctg cttccctgct gttttgtgga
     2761 atatctaccg actggaaaca ggcaaatgca ggaaattact gaactgaggg gacaggcgag
     2821 agacgatgcc aaagagctac accgacgagc tggccgagtg ggttgaatcc cgcgcggcca
     2881 agaagcgccg gcgtgatgag gctgcggttg cgttcctggc ggtgagggcg gatgtcgagg
     2941 cggcgttagc gtccggctat gcgctcgtca ccatttggga gcacatgcgg gaaacgggga
     3001 aggtcaagtt ctcctacgag acgttccgct cgcacgccag gcggcacatc aaggccaagc
     3061 ccgccgatgt gcccgcaccg caggccaagg ctgcggaacc cgcgccggca cccaagacgc
     3121 cggagccacg gcggccgaag caggggggca aggctgaaaa gccggccccc gctgcggccc
     3181 cgaccggctt caccttcaac ccaacaccgg acaaaaagga tcctctacgc cggacgcatc
     3241 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
     3301 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
     3361 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgctgcc tcgcgcgttt
     3421 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     3481 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     3541 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     3601 gcggcatcag agcagattgt actgagagtg caccataaat caatctaaag tatatatgag
     3661 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3721 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3781 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3841 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3901 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3961 gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg
     4021 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4081 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4141 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4201 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4261 atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc
     4321 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4381 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4441 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4501 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4561 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4621 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgc agatctgcag
     4681 gtcgaccgag agctcgaaaa caaactaaag cgcccttgtg gcgctttagt tttgttagag
     4741 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttaatt tgatgcctgg
     4801 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcgca acgttcaaat
     4861 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     4921 gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctggca gttccctact
     4981 ctcgcatggg gagaccccac actaccatcg gcgctacggc gtttcacttc tgagttcggc
     5041 atggggtcag gtgggaccac cgcgctactg ccgccaggca aattctgttt tatcagaccg
     5101 cttctgcgtt ctgatttaat ctgtatcagg ctgaaaatct tctctcatcc gccaaaacag
     5161 cgagctctca gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca
     5221 gtttactttg cagggcttcc caaccttccc agagggcgcc ccagctggca attccggttc
     5281 gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac tgcaagctac
     5341 ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
     5401 ccggggtcag caccgtttct gcggactggc tttctacgtg ttccgcttcc tttagcagcc
     5461 cttgcgccct gagtgcttgc ggcagcgtga agctttctct gagctgtaac agcctgaccg
     5521 caacaaacga gaggatcgag accatccgct ccagattatc cggctcctcc atgcgttgcc
     5581 tctcggctcc tgctccggtt ttccatgcct tatggaactc ctcgatccgc cagcgatggg
     5641 tataaatgtc gatgacgcgc aaggcttggg ctagcgactc gaccggttcg ctggtcagca
     5701 acaaccattt caacggggtc tcacccttgg gcgggttaat ctcctcggcc agcaccgcgt
     5761 tgagcgtgat attcccctgt tttagcgtga tgcgcccact gcgcaggctc aagctcgcct
     5821 tgcgggctgg tcgattttta cgtttaccgc gtttatccac cacgcccttt tgcggaatgc
     5881 tgatctgata gccacccaac tccggttggt tcttcagatg gtcgtacaga tacaacccag
     5941 actctacgtc cttgcgtggg tgcttggagc gcaccacgaa gcgctcgtta tgcgccagtt
     6001 tgtcctgcag ataagcatga atatcggctt cgcggtcaca gaccgcaatc acgttgctca
     6061 tcatgctgcc catgcgtaac cggctagttg cggcggctgc cagccatttg ccactctcct
     6121 tttcatccgc atcggcaggg tcatccgggc gcatccacca ctcctgatgc agtaatccta
     6181 cggtgcggaa tgtggtggcc tcgagcaaga gaacggagtg aacccaccat ccgcgggatt
     6241 tatcctgaat agagcccagc ttgccaagct cttcggcgac ctggtggcga taactcaaag
     6301 aggtggtgtc ctcaatggcc agcagttcgg gaaactcctg agccaacttg actgtttgca
     6361 tggcgccagc ctttctgatc gcctcggcag aaacgttggg attgcggata aatcggtaag
     6421 cgccttcctg ggcggcttta ctaccctctg atgagatggt tattgattta ccagaatatt
     6481 ttgccaattg ggcggcgacg ttaaccaagc gggcagtacg gcgaggatca cccagcgccg
     6541 ccgaagagaa cacagattta gcccagtcgg ccgcacgatg aagagcagaa gttatcatat
     6601 gtttcctgtg tgaaattgtt atccgctcac aattccacac attatacgag ccgatgatta
     6661 attgtcaaca gctcatttca gaatatttgc cagaaccgtt atgatgtcgg cgcaaaaaac
     6721 attatccaga acgggagtgc gccttgagcg acacgaatta tgcagtgatt tacgacctgc
     6781 acagccatac cacagcttcc gatggctgcc tgacgccaga agcattggtg caccgtgcag
     6841 tcgataagct ggtaccgaac gttaccatgt taggaggtca catggaagat cagatcctgg
     6901 aaaacgggaa aggttccgtt caggacgcta cttgtgtacg ggatcggtcg acggatccca
     6961 agcttcttct aga
//
