LOCUS       40924_27119        7209 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Transposon delivery vector pKUT-Tn5-Km DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7209)
  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  JOURNAL   Unpublished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 7209)
  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 7209)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7209)
  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            "Unpublished"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..7209
                     /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(60..851)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    complement(1236..1254)
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     protein_bind    1266..1282
                     /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(1317..1705)
                     /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(2326..2694)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(2727..2836)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(3917..4774)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4775..4879)
                     /label=AmpR promoter
     terminator      complement(4933..4969)
                     /label=soxR terminator
                     /note="bidirectional E. coli soxR transcription terminator"
     terminator      complement(4991..5018)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(5110..5196)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(5407..6834)
                     /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(6852..6868)
                     /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(6876..6904)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
ORIGIN
        1 ctgtctctta tacacatcta cgcgtcgaat taattccgcg aaccccagag tcccgctcag
       61 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
      121 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
      181 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
      241 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg
      301 agatcctcgc cgtcgggcat ccgcgccttg agcctggcga acagttcggc tggcgcgagc
      361 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
      421 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
      481 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
      541 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
      601 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
      661 gcctcgtctt ggagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
      721 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
      781 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
      841 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cagatcttga tcccctgcgc
      901 catcagatcc ttggcggcaa gaaagccatc cagtttactt tgcagggctt cccaacctta
      961 ccagagggcg ccccagctgg caattccggt tcgcttgctg tccataaaac cgcccagtct
     1021 agctatcgcc atgtaagccc actgcaagct acctgctttc tctttgcgct tgcgttttcc
     1081 cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg
     1141 gctttctacg tgttccgctt cctttagcag cccttgcgcc ctgagtgctt gcggcagcgt
     1201 gaagctagcg gaattaattc gacgcgtgcg gccgcagatg tgtataagag acagaattcc
     1261 acaaattgtt atccgctcac aattccacat gtggaattcc acatgtggaa ttcccatgtc
     1321 agccgttaag tgttcctgtg tcactcaaaa ttgctttgag aggctctaag ggcttctcag
     1381 tgcgttacat ccctggcttg ttgtccacaa ccgttaaacc ttaaaagctt taaaagcctt
     1441 atatattctt ttttttctta taaaacttaa aaccttagag gctatttaag ttgctgattt
     1501 atattaattt tattgttcaa acatgagagc ttagtacgtg aaacatgaga gcttagtacg
     1561 ttagccatga gagcttagta cgttagccat gagggtttag ttcgttaaac atgagagctt
     1621 agtacgttaa acatgagagc ttagtacgtg aaacatgaga gcttagtacg tactatcaac
     1681 aggttgaact gctgatcttc agatcctcta cgccggacgc atcgtggccg gatccagccg
     1741 accaggcttt ccacgcccgc gtgccgctcc atgtcgttcg cgcggttctc ggaaacgcgc
     1801 tgccgcgttt cgtgattgtc acgctcaagc ccgtagtccc gttcgagcgt cgcgcagagg
     1861 tcagcgaggg cgcggtaggc ccgatacggc tcatggatgg tgtttcgggt cgggtgaatc
     1921 ttgttgatgg cgatatggat gtgcaggttg tcggtgtcgt gatgcacggc actgacgcgc
     1981 tgatgctcgg cgaagccaag cccagcgcag atgcggtcct caatcgcgcg caacgtctcc
     2041 gcgtcgggct tctctcccgc gcggaagcta accagcaggt gataggtctt gtcggcctcg
     2101 gaacgggtgt tgccgtgctg ggtcgccatc acctcggcca tgacagcggg cagggtgttt
     2161 gcctcgcagt tcgtgacgcg cacgtgaccc aggcgctcgg tcttgccttg ctcgtcggtg
     2221 atgtacttca ccagctccgc gaagtcgctc ttcttgatgg agcgcatggg gacgtgcttg
     2281 gcaatcacgc gcaccccccg gccgttttag cggctaaaaa agtcatggct ctgccctcgg
     2341 gcggaccacg cccatcatga ccttgccaag ctcgtcctgc ttctcttcga tcttcgccag
     2401 cagggcgagg atcgtggcat caccgaaccg cgccgtgcgc gggtcgtcgg tgagccagag
     2461 tttcagcagg ccgcccaggc ggcccaggtc gccattgatg cgggccagct cgcggacgtg
     2521 ctcatagtcc acgacgcccg tgattttgta gccctggccg acggccagca ggtaggccga
     2581 caggctcatg ccggccgccg ccgccttttc ctcaatcgct cttcgttcgt ctggaaggca
     2641 gtacaccttg ataggtgggc tgcccttcct ggttggcttg gtttcatcag ccatccgctt
     2701 gccctcatct gttacgccgg cggtagccgg ccagcctcgc agagcaggat tcccgttgag
     2761 caccgccagg tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac
     2821 ttcacctatc ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct
     2881 ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat
     2941 gaccccgaag cagggttatg cagcggaaaa gcgctgcttc cctgctgttt tgtggaatat
     3001 ctaccgactg gaaacaggca aatgcaggaa attactgaac tgaggggaca ggcgagagac
     3061 gatgccaaag agctacaccg acgagctggc cgagtgggtt gaatcccgcg cggccaagaa
     3121 gcgccggcgt gatgaggctg cggttgcgtt cctggcggtg agggcggatg tcgaggcggc
     3181 gttagcgtcc ggctatgcgc tcgtcaccat ttgggagcac atgcgggaaa cggggaaggt
     3241 caagttctcc tacgagacgt tccgctcgca cgccaggcgg cacatcaagg ccaagcccgc
     3301 cgatgtgccc gcaccgcagg ccaaggctgc ggaacccgcg ccggcaccca agacgccgga
     3361 gccacggcgg ccgaagcagg ggggcaaggc tgaaaagccg gcccccgctg cggccccgac
     3421 cggcttcacc ttcaacccaa caccggacaa aaaggatcct ctacgccgga cgcatcgtgg
     3481 ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg
     3541 gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg
     3601 caggccccgt ggccggggga ctgttgggcg ccatctcctt gctgcctcgc gcgtttcggt
     3661 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     3721 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     3781 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
     3841 catcagagca gattgtactg agagtgcacc ataaatcaat ctaaagtata tatgagtaaa
     3901 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3961 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4021 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4081 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4141 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4201 atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg
     4261 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4321 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4381 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4441 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4501 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     4561 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4621 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4681 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4741 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4801 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4861 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgcagat ctgcaggtcg
     4921 accgagagct cgaaaacaaa ctaaagcgcc cttgtggcgc tttagttttg ttagagtttg
     4981 tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat gcctggcagt
     5041 ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt tcaaatccgc
     5101 tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
     5161 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg
     5221 catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag ttcggcatgg
     5281 ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc agaccgcttc
     5341 tgcgttctga tttaatctgt atcaggctga aaatcttctc tcatccgcca aaacagcgag
     5401 ctctcagatc ttgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt
     5461 actttgcagg gcttcccaac cttcccagag ggcgccccag ctggcaattc cggttcgctt
     5521 gctgtccata aaaccgccca gtctagctat cgccatgtaa gcccactgca agctacctgc
     5581 tttctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccgg
     5641 ggtcagcacc gtttctgcgg actggctttc tacgtgttcc gcttccttta gcagcccttg
     5701 cgccctgagt gcttgcggca gcgtgaagct ttctctgagc tgtaacagcc tgaccgcaac
     5761 aaacgagagg atcgagacca tccgctccag attatccggc tcctccatgc gttgcctctc
     5821 ggctcctgct ccggttttcc atgccttatg gaactcctcg atccgccagc gatgggtata
     5881 aatgtcgatg acgcgcaagg cttgggctag cgactcgacc ggttcgctgg tcagcaacaa
     5941 ccatttcaac ggggtctcac ccttgggcgg gttaatctcc tcggccagca ccgcgttgag
     6001 cgtgatattc ccctgtttta gcgtgatgcg cccactgcgc aggctcaagc tcgccttgcg
     6061 ggctggtcga tttttacgtt taccgcgttt atccaccacg cccttttgcg gaatgctgat
     6121 ctgatagcca cccaactccg gttggttctt cagatggtcg tacagataca acccagactc
     6181 tacgtccttg cgtgggtgct tggagcgcac cacgaagcgc tcgttatgcg ccagtttgtc
     6241 ctgcagataa gcatgaatat cggcttcgcg gtcacagacc gcaatcacgt tgctcatcat
     6301 gctgcccatg cgtaaccggc tagttgcggc ggctgccagc catttgccac tctccttttc
     6361 atccgcatcg gcagggtcat ccgggcgcat ccaccactcc tgatgcagta atcctacggt
     6421 gcggaatgtg gtggcctcga gcaagagaac ggagtgaacc caccatccgc gggatttatc
     6481 ctgaatagag cccagcttgc caagctcttc ggcgacctgg tggcgataac tcaaagaggt
     6541 ggtgtcctca atggccagca gttcgggaaa ctcctgagcc aacttgactg tttgcatggc
     6601 gccagccttt ctgatcgcct cggcagaaac gttgggattg cggataaatc ggtaagcgcc
     6661 ttcctgggcg gctttactac cctctgatga gatggttatt gatttaccag aatattttgc
     6721 caattgggcg gcgacgttaa ccaagcgggc agtacggcga ggatcaccca gcgccgccga
     6781 agagaacaca gatttagccc agtcggccgc acgatgaaga gcagaagtta tcatatgttt
     6841 cctgtgtgaa attgttatcc gctcacaatt ccacacatta tacgagccga tgattaattg
     6901 tcaacagctc atttcagaat atttgccaga accgttatga tgtcggcgca aaaaacatta
     6961 tccagaacgg gagtgcgcct tgagcgacac gaattatgca gtgatttacg acctgcacag
     7021 ccataccaca gcttccgatg gctgcctgac gccagaagca ttggtgcacc gtgcagtcga
     7081 taagctggta ccgaacgtta ccatgttagg aggtcacatg gaagatcaga tcctggaaaa
     7141 cgggaaaggt tccgttcagg acgctacttg tgtacgggat cggtcgacgg atcccaagct
     7201 tcttctaga
//
