LOCUS       40924_19901       10504 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Conjugative cloning vector pFE604T DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10504)
  AUTHORS   Yong HT, Yamamoto N, Takeuchi R, Hsieh YJ, Conrad TM, Datsenko KA, 
            Nakayashiki T, Wanner BL, Mori H.
  TITLE     Development of a System for Discovery of Genetic Interactions for 
            Essential Genes in Escherichia coli K-12
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10504)
  AUTHORS   Mori H, Yong HT.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-AUG-2013) Contact:Hirotada Mori Nara Institute of 
            Science and Technology, Graduate School of Biological Sciences; 
            8916-5 Takayama, Ikoma, Nara 630-0101, Japan URL 
            :http://ecoli.naist.jp/Lab
REFERENCE   3  (bases 1 to 10504)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10504)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-AUG-2013) Contact:Hirotada Mori Nara Institute of Science and 
            Technology, Graduate School of Biological Sciences; 8916-5 Takayama,
            Ikoma, Nara 630-0101, Japan URL :http://ecoli.naist.jp/Lab"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..67
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     protein_bind    44..60
                     /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)."
     protein_bind    complement(74..93)
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     RBS             110..121
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      502..596
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        933..961
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1131..1661
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     oriT            1684..2018
     rep_origin      complement(2037..2425)
                     /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(2489..3112)
                     /label=TetR
                     /note="tetracycline repressor TetR"
     promoter        3128..3183
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     CDS             3191..4393
                     /label=TcR
                     /note="tetracycline efflux protein"
     misc_feature    complement(4851..5324)
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(5400..6368)
                     /label=sopB
                     /note="partitioning protein for the bacterial F plasmid"
     CDS             complement(6371..7543)
                     /label=sopA
                     /note="partitioning protein for the bacterial F plasmid"
     misc_feature    complement(7869..8119)
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(8125..8877)
                     /label=repE
                     /note="replication initiation protein for the bacterial F 
                     plasmid"
     rep_origin      complement(8968..9187)
                     /direction=LEFT
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     promoter        9331..9408
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             9409..10488
                     /label=lacI
                     /note="lac repressor"
ORIGIN
        1 cctttcgtct tcacctcgag aaatcataaa aaatttattt gctttgtgag cggataacaa
       61 ttataatgtg tgaaattgtg agcgctcaca atttcacaca gaattcatta aagaggagaa
      121 attaactatg gccctgaggg ccgcagaggg atgaagtcga tcgcaccgtt acacatatgc
      181 aggatgaagc agccaacttc ccggacccgg tagaccgtgc agcccaggaa gaagagttca
      241 gcctcgaact gcgtaaccgc gatcgcgagc gtaagctgat caaaaagatc gagaagacgc
      301 tgaaaaaagt ggaagacgaa gatttcggct actgcgaatc ctgcggtgtt gaaattggta
      361 ttcgccgtct ggaagcgcgc ccgacagccg atctgtgcat cgactgcaaa acgctggctg
      421 aaattcgcga aaaacagatg gctggcggcc tatgcggccg ctaagggtcg acctgcagcc
      481 aagcttaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
      541 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccaa
      601 gctaggggaa ttcatcgtga ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt
      661 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
      721 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
      781 atgacccagt cacgtagcga tagcggagtg tataattctt gaagacgaaa gggcctcgtg
      841 atacgctcgc acgggcccat ctggcagttc cctactctcg catggggaga ccccacacta
      901 ccatcggggg ccacaggcgc ccacgaaccc agttgacata agcctgttcg gttcgtaaac
      961 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     1021 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt gtacagtcta
     1081 tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga
     1141 gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaggt ggctcaagta
     1201 tgggcatcat tcgcacatgt aggctcggcc ctgaccaagt caaatccatg cgggctgctc
     1261 ttgatctttt cggtcgtgag ttcggagacg tagccaccta ctcccaacat cagccggact
     1321 ccgattacct cgggaacttg ctccgtagta agacattcat cgcgcttgct gccttcgacc
     1381 aagaagcggt tgttggcgct ctcgcggctt acgttctgcc caagtttgag cagccgcgta
     1441 gtgagatcta tatctatgat ctcgcagtct ccggagagca ccggaggcag ggcattgcca
     1501 ccgcgctcat caatctcctc aagcatgagg ccaacgcgct tggtgcttat gtgatctacg
     1561 tgcaagcaga ttacggtgac gatcccgcag tggctctcta tacaaagttg ggcatacggg
     1621 aagaagtgat gcactttgat atcgacccaa gtaccgccac ctaacaattc gttcaaggcg
     1681 gccgccacct ctggtgactt tatccgtaaa taatttaacc cactccacaa aaaggctcaa
     1741 caggttggtg gttctcacca ccaaaagcac cacaccccac gcaaaaacaa gtttttgctg
     1801 atttttcttt ataaatagag tgttatgaaa aattagtttc tcttactctc tttatgatat
     1861 ttaaaaaagc ggtgtcggcg cggctacaac aacgcgccga caccgttttg taggggtggt
     1921 actgactatt tttataaaaa acattatttt atattagggg tgctgctagc ggcgcggtgt
     1981 gtttttttat aggataccgc taggggcgct gcgcggccgc aagaggctaa ttcccatgtc
     2041 agccgttaag tgttcctgtg tcactgaaaa ttgctttgag aggctctaag ggcttctcag
     2101 tgcgttacat ccctggcttg ttgtccacaa ccgttaaacc ttaaaagctt taaaagcctt
     2161 atatattctt ttttttctta taaaacttaa aaccttagag gctatttaag ttgctgattt
     2221 atattaattt tattgttcaa acatgagagc ttagtacgtg aaacatgaga gcttagtacg
     2281 ttagccatga gagcttagta cgttagccat gagggtttag ttcgttaaac atgagagctt
     2341 agtacgttaa acatgagagc ttagtacgtg aaacatgaga gcttagtacg tactatcaac
     2401 aggttgaact gctgatcttc agatcctcta cgccggacgc ttggttaccg tgaagttacc
     2461 atcacggaaa aaggttatgc tgcttttaag acccactttc acatttaagt tgtttttcta
     2521 atccgcatat gatcaattca aggccgaata agaaggctgg ctctgcacct tggtgatcaa
     2581 ataattcgat agcttgtcgt aataatggcg gcatactatc agtagtaggt gtttcccttt
     2641 cttctttagc gacttgatgc tcttgatctt ccaatacgca acctaaagta aaatgcccca
     2701 cagcgctgag tgcatataat gcattctcta gtgaaaaacc ttgttggcat aaaaaggcta
     2761 attgattttc gagagtttca tactgttttt ctgtaggccg tgtacctaaa tgtacttttg
     2821 ctccatcgcg atgacttagt aaagcacatc taaaactttt agcgttatta cgtaaaaaat
     2881 cttgccagct ttccccttct aaagggcaaa agtgagtatg gtgcctatct aacatctcaa
     2941 tggctaaggc gtcgagcaaa gcccgcttat tttttacatg ccaatacaat gtaggctgct
     3001 ctacacctag cttctgggcg agtttacggg ttgttaaacc ttcgattccg acctcattaa
     3061 gcagctctaa tgcgctgtta atcactttac ttttatctaa tctagacatc attaattcct
     3121 aatttttgtt gacactctat cattgataga gttattttac cactccctat cagtgataga
     3181 gaaaagtgaa atgaatagtt cgacaaagat cgcattggta attacgttac tcgatgccat
     3241 ggggattggc cttatcatgc cagtcttgcc aacgttatta cgtgaattta ttgcttcgga
     3301 agatatcgct aaccactttg gcgtattgct tgcactttat gcgttaatgc aggttatctt
     3361 tgctccttgg cttggaaaaa tgtctgaccg atttggtcgg cgcccagtgc tgttgttgtc
     3421 attaataggc gcatcgctgg attacttatt gctggctttt tcaagtgcgc tttggatgct
     3481 gtatttaggc cgtttgcttt cagggatcac aggagctact ggggctgtcg cggcatcggt
     3541 cattgccgat accacctcag cttctcaacg cgtgaagtgg ttcggttggt taggggcaag
     3601 ttttgggctt ggtttaatag cggggcctat tattggtggt tttgcaggag agatttcacc
     3661 gcatagtccc ttttttatcg ctgcgttgct aaatattgtc actttccttg tggttatgtt
     3721 ttggttccgt gaaaccaaaa atacacgtga taatacagat accgaagtag gggttgagac
     3781 gcaatcgaat tcggtataca tcactttatt taaaacgatg cccattttgt tgattattta
     3841 tttttcagcg caattgatag gccaaattcc cgcaacggtg tgggtgctat ttaccgaaaa
     3901 tcgttttgga tggaatagca tgatggttgg cttttcatta gcgggtcttg gtcttttaca
     3961 ctcagtattc caagcctttg tggcaggaag aatagccact aaatggggcg aaaaaacggc
     4021 agtactgctc gaatttattg cagatagtag tgcatttgcc tttttagcgt ttatatctga
     4081 aggttggtta gatttccctg ttttaatttt attggctggt ggtgggatcg ctttacctgc
     4141 attacaggga gtgatgtcta tccaaacaaa gagtcatgag caaggtgctt tacagggatt
     4201 attggtgagc cttaccaatg caaccggtgt tattggccca ttactgttta ctgttattta
     4261 taatcattca ctaccaattt gggatggctg gatttggatt attggtttag cgttttactg
     4321 tattattatc ctgctatcga tgaccttcat gttaacccct caagctcagg ggagtaaaca
     4381 ggagacaagt gcttagttat ttcgtcacca aatgatgtta ttccgcgaaa tattacgcct
     4441 gaataagtga taataagcgg atgaatggca gaaattctcc tctcacatgc ggcctccatc
     4501 aggccgtgcc gcacctgtaa ccgggcgacc gcatacgaat gcaggtgcgg ccccgaagcc
     4561 gagcgcgtgg aggagaaacg acgaaacgtg accgggtgag gaggccgtcg acagcgacac
     4621 acttgcatcg gatgcagccc ggttaacgtg ccggcacggc ctgggtaacc aggtattttg
     4681 tccacataac cgtgcgcaaa atgttgtgga taagcaggac acagcagcaa tccacagcag
     4741 gcatacaacc gcacaccgag gttactccgt tctacaggtt acgacgacat gtcaatactt
     4801 gcccttgaca ggcattgatg gaatcgtagt ctcacgctga tagtctgatc gacaatacaa
     4861 gtgggaccgt ggtcccagac cgataatcag accgacaaca cgagtgggat cgtggtccca
     4921 gactaataat cagaccgacg atacgagtgg gaccgtggtc ccagactaat aatcagaccg
     4981 acgatacgag tgggaccgtg gttccagact aataatcaga ccgacgatac gagtgggacc
     5041 gtggtcccag actaataatc agaccgacga tacgagtggg accatggtcc cagactaata
     5101 atcagaccga cgatacgagt gggaccgtgg tcccagtctg attatcagac cgacgatacg
     5161 agtgggaccg tggtcccaga ctaataatca gaccgacgat acgagtggga ccgtggtccc
     5221 agactaataa tcagaccgac gatacgagtg ggaccgtggt cccagtctga ttatcagacc
     5281 gacgatacaa gtggaacagt gggcccagag agaatattca ggccagttat gctttctggc
     5341 ctgtaacaaa ggacattaag taaagacaga taaacgtaga ctaaaacgtg gtcgcatcag
     5401 ggtgctggct tttcaagttc cttaagaatg gcctcaattt tctctataca ctcagttgga
     5461 acacgagacc tgtccaggtt aagcaccatt ttatcgccct tatacaatac tgtcgctcca
     5521 ggagcaaact gatgtcgtga gcttaaacta gttcttgatg cagatgacgt tttaagcaca
     5581 gaagttaaaa gagtgataac ttcttcagct tcaaatatca ccccagcttt tttctgctca
     5641 tgaaggttag atgcctgctg cttaagtaat tcctctttat ctgtaaaggc tttttgaagt
     5701 gcatcacctg accgggcaga tagttcaccg gggtgagaaa aaagagcaac aactgattta
     5761 ggcaatttgg cggtgttgat acagcgggta ataatcttac gtgaaatatt ttccgcatca
     5821 gccagcgcag aaatatttcc agcaaattca ttctgcaatc ggcttgcata acgctgacca
     5881 cgttcataag cacttgttgg gcgataatcg ttacccaatc tggataatgc agccatctgc
     5941 tcatcatcca gctcgccaac cagaacacga taatcacttt cggtaagtgc agcagcttta
     6001 cgacggcgac tcccatcggc aatttctatg acaccagata ctcttcgacc gaacgccggt
     6061 gtctgttgac cagtcagtag aaaagaaggg atgagatcat ccagtgcgtc ctcagtaagc
     6121 agctcctggt cacgttcatt acctgaccat acccgagagg tcttctcaac actatcaccc
     6181 cggagcactt caagagtaaa cttcacatcc cgaccacata caggcaaagt aatggcatta
     6241 ccgcgagcca ttactcctac gcgcgcaatt aacgaatcca ccatcggggc agctggtgtc
     6301 gataacgaag tatcttcaac cggttgagta ttgagcgtat gttttggaat aacaggcgca
     6361 cgcttcatta tctaatctcc cagcgtggtt taatcagacg atcgaaaatt tcattgcaga
     6421 caggttccca aatagaaaga gcatttctcc aggcaccagt tgaagagcgt tgatcaatgg
     6481 cctgttcaaa aacagttctc atccggatct gacctttacc aacttcatcc gtttcacgta
     6541 caacattttt tagaaccatg cttccccagg catcccgaat ttgctcctcc atccacgggg
     6601 actgagagcc attactattg ctgtatttgg taagcaaaat acgtacatca ggctcgaacc
     6661 ctttaagatc aacgttcttg agcagatcac gaagcatatc gaaaaactgc agtgcggagg
     6721 tgtagtcaaa caactcagca ggcgtgggaa caatcagcac atcagcagca catacgacat
     6781 taatcgtgcc gatacccagg ttaggcgcgc tgtcaataac tatgacatca tagtcatgag
     6841 caacagtttc aatggccagt cggagcatca ggtgtggatc ggtgggcagt ttaccttcat
     6901 caaatttgcc cattaactca gtttcaatac ggtgcagagc cagacaggaa ggaataatgt
     6961 caagccccgg ccagcaagtg ggctttattg cataagtgac atcgtccttt tccccaagat
     7021 agaaaggcag gagagtgtct tctgcatgaa tatgaagatc tggtacccat ccgtgataca
     7081 ttgaggctgt tccctggggg tcgttacctt ccacgagcaa aacacgtagc cccttcagag
     7141 ccagatcctg agcaagatga acagaaactg aggttttgta aacgccacct ttatgggcag
     7201 caaccccgat caccggtgga aatacgtctt cagcacgtcg caatcgcgta ccaaacacat
     7261 cacgcatatg attaatttgt tcaattgtat aaccaacacg ttgctcaacc cgtcctcgaa
     7321 tttccatatc cgggtgcggt agtcgccctg ctttctcggc atctctgata gcctgagaag
     7381 aaaccccaac taaatccgct gcttcaccta ttctccagcg ccgggttatt ttcctcgctt
     7441 ccgggctgtc atcattaaac tgtgcaatgg cgatagcctt cgtcatttca tgaccagcgt
     7501 ttatgcactg gttaagtgtt tccatgagtt tcattctgaa catcctttaa tcattgcttt
     7561 gcgttttttt attaaatctt gcaatttact gcaaagcaac aacaaaatcg caaagtcatc
     7621 aaaaaaccgc aaagttgttt aaaataagag caacactaca aaaggagata agaagagcac
     7681 atacctcagt cacttattat cactagcgct cgccgcagcc gtgtaaccga gcatagcgag
     7741 cgaactggcg aggaagcaaa gaagaactgt tctgtcagat agctcttacg ctcagcgcaa
     7801 gaagaaatat ccaccgtggg aaaaactcca ggtagaggta cacacgcgga tagccaattc
     7861 agagtaataa actgtgataa tcaaccctca tcaatgatga cgaactaacc cccgatatca
     7921 ggtcacatga cgaagggaaa gagaaggaaa tcaactgtga caaactgccc tcaaatttgg
     7981 cttccttaaa aattacagtt caaaaagtat gagaaaatcc atgcaggctg aaggaaacag
     8041 caaaactgtg acaaattacc ctcagtaggt cagaacaaat gtgacgaacc accctcaaat
     8101 ctgtgacaga taaccctcag actatcctgt cgtcatggaa gtgatatcgc ggaaggaaaa
     8161 tacgatatga gtcgtctggc ggcctttctt tttctcaatg tatgagaggc gcattggagt
     8221 tctgctgttg atctcattaa cacagacctg caggaagcgg cggcggaagt caggcatacg
     8281 ctggtaactt tgaggcagct ggtaacgctc tatgatccag tcgattttca gagagacgat
     8341 gcctgagcca tccggcttac gatactgaca cagggattcg tataaacgca tggcatacgg
     8401 attggtgatt tcttttgttt cactaagccg aaactgcgta aaccggttct gtaacccgat
     8461 aaagaaggga atgagatatg ggttgatatg tacactgtaa agccctctgg atggactgtg
     8521 cgcacgtttg ataaaccaag gaaaagattc atagcctttt tcatcgccgg catcctcttc
     8581 agggcgataa aaaaccactt ccttccccgc gaaactcttc aatgcctgcc gtatatcctt
     8641 actggcttcc gcagaggtca atccgaatat ttcagcatat ttagcaacat ggatctcgca
     8701 gataccgtca tgttcctgta gggtgccatc agattttctg atctggtcaa cgaacagata
     8761 cagcatacgt ttttgatccc gggagagact atatgccgcc tcagtgaggt cgtttgactg
     8821 gacgattcgc gggctatttt tacgtttctt gtgattgata accgctgttt ccgccatgac
     8881 agatccatgt gaagtgtgac aagtttttag attgtcacac taaataaaaa agagtcaata
     8941 agcagggata actttgtgaa aaaacagctt cttctgaggg caatttgtca cagggttaag
     9001 ggcaatttgt cacagacagg actgtcattt gagggtgatt tgtcacactg aaagggcaat
     9061 ttgtcacaac accttctcta gaaccagcat ggataaaggc ctacaaggcg ctctaaaaaa
     9121 gaagatctaa aaactataaa aaaaataatt ataaaaatat ccccgtggat aagtggataa
     9181 ccccaaggga agttttttca ggcatcgtgt gtaagcagaa tatataagtg ctgttccctg
     9241 gtgcttcctc gctcactcga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     9301 ctgtgcggta tttcacaccg cataaattcc gacaccatcg aatggtgcaa aacctttcgc
     9361 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta
     9421 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
     9481 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
     9541 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt
     9601 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa
     9661 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc
     9721 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt
     9781 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg
     9841 gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa
     9901 gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg
     9961 ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat
    10021 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc
    10081 ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt
    10141 gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt
    10201 ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg
    10261 ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg
    10321 ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg
    10381 aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
    10441 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg ataacggacc
    10501 gcgg
//
