LOCUS       V009018                 9952 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009018
VERSION     V009018
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9952)
  AUTHORS   Kang Y, Son MS, Hoang TT.
  TITLE     One step engineering of T7-expression strains for protein
            production: increasing the host-range of the T7-expression system
  JOURNAL   Protein Expr. Purif. 55 (2), 325-333 (2007)
   PUBMED   17716915
REFERENCE   2  (bases 1 to 9952)
  AUTHORS   Kang Y, Son MS, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-NOV-2006) Microbiology, University of Hawaii at Manoa,
            2538 The Mall - Snyder 310, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 9952)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9952)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein
            Expr. Purif."; date: "2007"; volume: "55"; issue: "2"; pages:
            "325-333"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (30-NOV-2006) Microbiology, University of Hawaii at Manoa, 2538 The
            Mall - Snyder 310, Honolulu, HI 96822, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             34..402
                     /label="traJ"
                     /note="oriT-recognizing protein"
     rep_origin      1021..1409
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     repeat_region   1451..1478
                     /rpt_family="Mariner"
     CDS             complement(1549..4197)
                     /note="T7 RNA polymerase from Escherichia phage T7.
                     Accession#: P00573"
                     /label="T7 RNA polymerase"
     CDS             complement(4506..4679)
                     /label="lacZ-alpha"
                     /note="LacZ-alpha fragment of beta-galactosidase"
     misc_feature    complement(4697..4721)
                     /label="lac operator"
                     /note="lac operator"
     protein_bind    4699..4715
                     /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)."
     promoter        complement(4723..4753)
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    complement(4768..4789)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4805..5884)
                     /label="lacI"
                     /note="lac repressor"
     misc_feature    complement(5965..6012)
                     /note="FRT; Flp-recombination-target"
     protein_bind    complement(5965..6012)
                     /label="FRT"
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        6053..6081
                     /label="Pc promoter"
                     /note="class 1 integron promoter"
     CDS             6270..6800
                     /label="GmR"
                     /note="gentamycin acetyltransferase"
     protein_bind    complement(6933..6980)
                     /label="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             7243..7695
                     /label="T7 lysozyme"
                     /note="lysozyme from bacteriophage T7"
     repeat_region   7730..7757
                     /rpt_family="Mariner"
     primer_bind     7890..7906
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        7916..7934
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(7997..9043)
                     /codon_start=1
                     /gene="Tnp"
                     /product="Tnp"
                     /label="Tnp"
                     /note="mariner transposase"
                     /protein_id="ABN54776.1"
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDLKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     gene            complement(7997..9043)
                     /gene="Tnp"
                     /label="Tnp"
     oriT            9844..9952
                     /label="oriT"
                     /note="incP origin of transfer"
ORIGIN
        1 gctaccgccg gcgtaacaga tgagggcaag cggatggctg atgaaaccaa gccaaccagg
       61 aagggcagcc cacctatcaa ggtgtactgc cttccagacg aacgaagagc gattgaggaa
      121 aaggcggcgg cggccggcat gagcctgtcg gcctacctgc tggccgtcgg ccagggctac
      181 aaaatcacgg gcgtcgtgga ctatgagcac gtccgcgagc tggcccgcat caatggcgac
      241 ctgggccgcc tgggcggcct gctgaaactc tggctcaccg acgacccgcg cacggcgcgg
      301 ttcggtgatg ccacgatcct cgccctgctg gcgaagatcg aagagaagca ggacgagctt
      361 ggcaaggtca tgatgggcgt ggtccgcccg agggcagagc catgactttt ttagccgcta
      421 aaacggccgg ggggtgcgcg tgattgccaa gcacgtcccc atgcgctcca tcaagaagag
      481 cgacttcgcg gagctggtga agtacatcac cgacgagcaa ggcaagaccg agcgcctggg
      541 tcacgtgcgc gtcacgaact gcgaggcaaa caccctgccc gctgtcatgg ccgaggtgat
      601 ggcgacccag cacggcaaca cccgttccga ggccgacaag acctatcacc tgctggttag
      661 cttccgcgcg ggagagaagc ccgacgcgga gacgttgcgc gcgattgagg accgcatctg
      721 cgctgggctt ggcttcgccg agcatcagcg cgtcagtgcc gtgcatcacg acaccgacaa
      781 cctgcacatc catatcgcca tcaacaagat tcacccgacc cgaaacacca tccatgagcc
      841 gtatcgggcc taccgcgccc tcgctgacct ctgcgcgacg ctcgaacggg actacgggct
      901 tgagcgtgac aatcacgaaa cgcggcagcg cgtttccgag aaccgcgcga acgacatgga
      961 gcggcacgcg ggcgtggaaa gcctggtcgg ctggatcccc acgatgcgtc cggcgtagag
     1021 gatctgaaga tcagcagttc aacctgttga tagtacgtac taagctctca tgtttcacgt
     1081 actaagctct catgtttaac gtactaagct ctcatgttta acgaactaaa ccctcatggc
     1141 taacgtacta agctctcatg gctaacgtac taagctctca tgtttcacgt actaagcttt
     1201 catgtttgac caataaaatt aatataaatc agcaacttaa atagcttcta aggttttaag
     1261 ttttataaga aaaaaaagaa tatataaggc ttttaaagct tttaaggttt aacggttgtg
     1321 gacaacaagc cagggatgta acgcactgag aagcccttag agcctctcaa agcaattttg
     1381 agtgacacag gaacacttaa cggctgacat gggaattgtg ccctgtacaa gaaagcaggg
     1441 tcccgggagt aacaggttgg ctgataagtc cccggtctct agtgtacaag aaagctgggt
     1501 gatcctctag agcttgcatg cctgcaggtc gactctagag gatccttacg cgaacgcgaa
     1561 gtccgactct aagatgtcac ggaggttcaa gttaccttta gccggaagtg ctggcatttt
     1621 gtccaattga gactcgtgca actggtcagc gaactggtcg tagaaatcag ccagtacatc
     1681 acaagactca tatgtgtcaa ccatagtttc gcgcactgct ttgaacaggt tcgcagcgtc
     1741 agccggaatg gtaccgaagg agtcgtgaat cagtgcaaaa gattcgattc cgtacttctc
     1801 gtgtgcccac actacagtct tacgaaggtg gctaccgtct tggctgtgta caaagttagg
     1861 agcgatacca gactcctgtt tgtgtgcatc aatctcgcta tctttgttgg tgttaatggt
     1921 aggctgtaag cggaactgac cgaggaacat caggttcaag cgcgtctgaa taggcttctt
     1981 gtattcctgc cacacaggga aaccatcagg agttacccaa tgcacagcgc aacgcttgcg
     2041 aagaatctct ccagtcttct tatctttgac ctcagcagcc agcagcttag cagcagactt
     2101 aagccagttc attgcttcaa ccgcagctac caccgtcacg ctcacagatt cccaaatcag
     2161 cttagccatg tatccagcag cctgattcgg ctgagtgaac atcagaccct tgccggaatc
     2221 aatagctggc tgaatggtat cttccagcac ttgttgacgg aagccgaact ctttggaccc
     2281 gtaagccagc gtcatgactg aacgcttagt cacactgcga gtaacaccgt aagccagcca
     2341 ttgaccagcc agtgccttag tgcccagctt gactttctca gagatttcac cagtgttctc
     2401 atcggtcacg gtaactactt cgttatcggt cccattgatt gcgtctgctt gtagaatctc
     2461 gttgactttc ttagcaacaa tcccgtagat gtcctgaacg gtttcactag gaagcaagtt
     2521 aaccgcgcga ccacctacct catctcggag catcgcggag aagtgctgga tgccagagca
     2581 agacccgtca aacgccagcg gaagggagca gttatagctc aggccgtggt gctgtacccc
     2641 agcgtactca aagcagaacg caaggaagca gaacggagaa tcttgctcag cccaccaagt
     2701 gttctccagt ggagacttag cgcaagccat gatgttctcg tggttttcct caatgaactt
     2761 gatgcgctca gggaacggaa ccttatcgac acccgcacag tttgcaccgt ggattttcag
     2821 ccagtagtaa ccttccttac cgattggttt acctttcgcc agcgtaagca gtcctttggt
     2881 catatcgtta ccttgcgggt tgaacattga cacagcgtaa acacgaccgc gccagtccat
     2941 gttgtaaggg aaccagatgg ccttatggtt agcaaactta ttggcttgct caagcatgaa
     3001 ctcaaggctg atacggcgag acttgcgagc cttgtccttg cggtacacag cagcggcagc
     3061 acgtttccac gcggtgagag cctcaggatt catgtcgatg tcttccggtt tcatcgggag
     3121 ttcttcacgc tcaatcgcag ggatgtcctc gaccggacaa tgcttccact tggtgattac
     3181 gttggcgacc gctaggactt tcttgttgat tttccatgcg gtgttttgcg caatgttaat
     3241 cgctttgtac acctcaggca tgtaaacgtc ttcgtagcgc atcagtgctt tcttactgtg
     3301 agtacgcacc agcgccagag gacgacgacc gttagcccaa tagccaccac cagtaatgcc
     3361 agtccacggc ttaggaggaa ctacgcaagg ttggaacatc ggagagatgc cagccagcgc
     3421 acctgcacgg gttgcgatag cctcagcgta ttcaggtgcg agttcgatag tctcagagtc
     3481 ttgacctact acgccagcat tttggcggtg taagctaacc attccggttg actcaatgag
     3541 catctcgatg cagcgtactc ctacatgaat agagtcttcc ttatgccacg aagaccacgc
     3601 ctcgccaccg agtagaccct tagagagcat gtcagcctcg acaacttgca taaatgcttt
     3661 cttgtagacg tgccctacgc gcttgttgag ttgttcctca acgtttttct tgaagtgctt
     3721 agcttcaagg tcacggatac gaccgaagcg agcctcgtcc tcaatggccc gaccgattgc
     3781 gcttgctaca gcctgaacgg ttgtattgtc agcactggtt aggcaagcca gagtggtctt
     3841 aatggtgatg tacgctacgg cttccggctt gatttcttgc aggaactgga aggctgtcgg
     3901 gcgcttgccg cgcttagctt tcacttcctc aaaccagtcg ttgatgcgtg caatcatctt
     3961 agggagtagg gtagtgatga gaggcttggc ggcagcgtta tccgcaacct caccagcttt
     4021 aagttgacgc tcaaacatct tgcggaagcg tgcttcaccc atctcgtaag actcatgctc
     4081 aagggccaac tgttcgcgag ctaaacgctc accgtaatgg tcagccagag tgttgaacgg
     4141 gatagcagcc agttcgatgt cagagaagtc gttcttagcg atgttaatcg tgttcattta
     4201 gtgcctcttc cagttagtaa atccgggatc agatcccgaa cgccatcaaa aataattcgc
     4261 gtctggcctt cctgtagcca gctttcatca acattaaatg tgagcgagta acaacccgtc
     4321 ggattctccg tgggaacaaa cggcggattg accgtaatgg gataggttac gttggtgtag
     4381 atgggcgcat cgtaaccgtg catctgccag tttgagggga cgacgacagt atcggcctca
     4441 ggaagatcgc actccagcca gctttccggc accgcttctg gtgccggaaa ccaggcaaag
     4501 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     4561 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     4621 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aatccgtaat catggtcata
     4681 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacattatac gagccggaag
     4741 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4801 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4861 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     4921 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     4981 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttgac ggcgggatat
     5041 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca
     5101 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     5161 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     5221 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     5281 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     5341 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     5401 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     5461 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     5521 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     5581 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     5641 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     5701 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     5761 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     5821 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     5881 tcacattcac caccctgaat tgactctctt ttttgcacca ttgtggatcc tctagagtcg
     5941 acgaattagc ttcaaaagcg ctctgaagtt cctatacttt ctagagaata ggaacttcgg
     6001 aataggaact tcaagatccc ctgattccct ttgtcaacag caatggatcg aattgacata
     6061 agcctgttcg gttcgtaaac tgtaatgcaa gtagcgtatg cgctcacgca actggtccag
     6121 aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga
     6181 ctgttttttt gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg
     6241 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagcaacgat gttacgcagc
     6301 agggcagtcg ccctaaaaca aagttaggtg gctcaagtat gggcatcatt cgcacatgta
     6361 ggctcggccc tgaccaagtc aaatccatgc gggctgctct tgatcttttc ggtcgtgagt
     6421 tcggagacgt agccacctac tcccaacatc agccggactc cgattacctc gggaacttgc
     6481 tccgtagtaa gacattcatc gcgcttgctg ccttcgacca agaagcggtt gttggcgctc
     6541 tcgcggctta cgttctgccc aggtttgagc agccgcgtag tgagatctat atctatgatc
     6601 tcgcagtctc cggcgagcac cggaggcagg gcattgccac cgcgctcatc aatctcctca
     6661 agcatgaggc caacgcgctt ggtgcttatg tgatctacgt gcaagcagat tacggtgacg
     6721 atcccgcagt ggctctctat acaaagttgg gcatacggga agaagtgatg cactttgata
     6781 tcgacccaag taccgccacc taacaattcg ttcaagccga gatcggcttc ccggccgcgg
     6841 agttgttcgg taaattgtca caacgccgcg gccaattcga tctagaattc cattgagtaa
     6901 gtttttaagc acatcagctt caaaagcgct ctgaagttcc tatactttct agagaatagg
     6961 aacttcggaa taggaacttc aagatcccca attcgtcgac ctgcaggcat gcaaggagat
     7021 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     7081 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     7141 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatccg
     7201 ggtccccttt gatagattaa aaaggaaagg aggaaagaaa taatggctcg tgtacagttt
     7261 aaacaacgtg aatctactga cgcaatcttt gttcactgct cggctaccaa gccaagtcag
     7321 aatgttggtg tccgtgagat tcgccagtgg cacaaagagc agggttggct cgatgtggga
     7381 taccacttta tcatcaagcg agacggtact gtggaggcag gacgagatga gatggctgta
     7441 ggctctcacg ctaagggtta caaccacaac tctatcggcg tctgccttgt tggtggtatc
     7501 gacgataaag gtaagttcga cgctaacttt acgccagccc aaatgcaatc ccttcgctca
     7561 ctgcttgtca cactgctggc taagtacgaa ggcgctggtc ttcgcgccca tcatgaggtg
     7621 gcgccgaagg cttgcccttc gttcgacctt aagcgttggt gggagaagaa cgaactggtc
     7681 acttctgacc gtggataatt aattgaactc actaaagggg taccgctaga gaccggggac
     7741 ttatcagcca acctgttact cccgggaccc agctttcttg tacagggcac cggtgcgggc
     7801 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     7861 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata
     7921 cgactcacta tagggcgaat tggaggattc cggtctaaca aagaaaaaca catttttttg
     7981 tcaaaattcg tttttattat tcaacatagt tcccttcaag agcgatacaa cgattataac
     8041 gaccttccaa ttttttgata ccattttggt agtactcctt cggttttgcc tcaaaatagg
     8101 cctcagtttc ggcgatcacc tcttcattgc agccaaattt tttccctgcg agcatccttt
     8161 tgaggtctga gaacaagaaa aagtcgctgg gggccagatc tggagaatac ggtgggtggg
     8221 gaagcaattc gaagcccaat tcatgaattt ttgccatcgt tctcaatgac ttgtggcacg
     8281 gtgcgttgtc ttggtggaac aacacttttt tcttcttcat gtggggccgt tttgccgcga
     8341 tttcgacctt caaacgctcc aataacgcca tataatagtc actgttgatg gtttttccct
     8401 tctcaagata atcgataaaa attattccat gcgcatccca aaaaacagag gccattactt
     8461 tgccagcgga cttttgagtc tttccacgct tcggagacgg ttcaccggtc gctgtccact
     8521 cagccgactg tcgattggac tcaggagtgt agtgatggag ccatgtttca tccattgtca
     8581 catatcgacg gaaaaactcg ggtgtattac gagttaacag ctgcaaacac cgcttagaat
     8641 catcaacacg tcgttgtttt tggtcaaatg tgagctcgcg cggcacccat ttcgcacaga
     8701 gcttccgcat atccaaatat tgatgaatga tatgaccaac acgttccttt gatatcttta
     8761 aggcctctgc tatctcgatc aacttcattt tacggtcatt caaaatcatt ttgtggattt
     8821 ttttgatgtt ttcgtcggta accacctctt tcgggcgtcc actgcgttca ccgtcctccg
     8881 tgctcatttc accacgcttg aattttgcat accaatcaat tattgttgat ttccctgggg
     8941 cagagtccgg aaactcatta tcaagccaag tttttgcttc cactgtattt tttcccttca
     9001 gaaaacagta ttttatcaaa acacgaaatt cctttttttc catatgtttc acaataacaa
     9061 aagttgcttc acaaaagacg ctctatctca caaactaatt gacttacaga cgtcaaattt
     9121 tgacacgaat catttgaagg ttggtactat ataaaaataa tatgcattta atactagcga
     9181 cgccatctat gtgtcaatcc atcacactgg cggccctagt ggatcctttt tgtccggtgt
     9241 tgggttgaag gtgaagccgg tcggggccgc agcgggggcc ggcttttcag ccttgccccc
     9301 ctgcttcggc cgccgtggct ccggcgtctt gggtgccggc gcgggttccg cagccttggc
     9361 ctgcggtgcg ggcacatcgg cgggcttggc cttgatgtgc cgcctggcgt gcgagcggaa
     9421 cgtctcgtag gagaacttga ccttccccgt ttcccgcatg tgctcccaaa tggtgacgag
     9481 cgcatagccg gacgctaacg ccgcctcgac atccgccctc accgccagga acgcaaccgc
     9541 agcctcatca cgccggcgct tcttggccgc gcgggattca acccactcgg ccagctcgtc
     9601 ggtgtagctc tttggcatcg tctctcgcct gtcccctcag ttcagtaatt tcctgcattt
     9661 gcctgtttcc agtcggtaga tattccacaa aacagcaggg aagcagcgct tttccgctgc
     9721 ataaccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg
     9781 atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca
     9841 gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc
     9901 ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cg
//
