LOCUS       V003840                 7163 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003840
VERSION     V003840
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7163)
  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 7163)
  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 7163)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7163)
  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..7163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(7..25)
                     /label="Tn5 ME"
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     protein_bind    37..268
                     /label="lambda attP"
                     /note="integrase from phage lambda"
     rep_origin      complement(561..946)
                     /direction=LEFT
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     misc_feature    951..969
                     /label="Tn5 ME"
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     CDS             complement(981..3629)
                     /note="T7 RNA polymerase from Escherichia phage T7.
                     Accession#: P00573"
                     /label="T7 RNA polymerase"
     CDS             complement(3938..4111)
                     /label="lacZ-alpha"
                     /note="LacZ-alpha fragment of beta-galactosidase"
     protein_bind    complement(4131..4147)
                     /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(4155..4185)
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    complement(4200..4221)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4237..5316)
                     /label="lacI"
                     /note="lac repressor"
     misc_feature    complement(5397..5444)
                     /label="Flp-recombination-target"
                     /note="Flp-recombination-target"
     protein_bind    complement(5397..5444)
                     /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        5485..5513
                     /label="Pc promoter"
                     /note="class 1 integron promoter"
     CDS             5702..6232
                     /label="GmR"
                     /note="gentamycin acetyltransferase"
     protein_bind    complement(6365..6412)
                     /label="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             6675..7127
                     /label="T7 lysozyme"
                     /note="lysozyme from bacteriophage T7"
ORIGIN
        1 caagctagat gtgtataaga gacagcccgg gaaatcaaat aatgatttta ttttgactga
       61 tagtgacctg ttcgttgcaa caaattgata agcaatgctt ttttataatg ccaacttagt
      121 ataaaaaagc tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt
      181 tgcataaaaa acagactaca taatactgta aaacacaaca tatgcagtca ctatgaatca
      241 actacttaga tggtattagt gacctgtaac agagcattag cgcaaggtgt ttttgtcttc
      301 ttgcgctaat tttttgtcat caaacctgtc gcactccaga gaagcacaaa gcctcgcaat
      361 ccagtgcaaa gctgaattgg ggatcattta ttctgcctcc cagagcctga taaaaacggt
      421 gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca
      481 ccgattattg aagcatttat cagggttatt gtctcatgac caaaatccct taacgtgagt
      541 tttcgttcca ctgagcgtca tgtcagccgt taagtgttcc tgtgtcactg aaaattgctt
      601 tgagaggctc taagggcttc tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta
      661 aaccttaaaa gctagcttta aaagccttat atattctttt ttttcttata aaacttaaaa
      721 ccttagaggc tatttaagtt gctgatttat attaatttta ttgttcaaac atgagagctt
      781 agtacgtgaa acatgagagc ttagtacgtt agccatgaga gcttagtacg ttagccatga
      841 gggtttagtt cgttaaacat gagagcttag tacgttaaac atgagagctt agtacgtgaa
      901 acatgagagc ttagtacgta ctatcaacag gttgaactgc tgatcccggg ctgtctctta
      961 tacacatctg gggatcctta cgcgaacgcg aagtccgact ctaagatgtc acggaggttc
     1021 aagttacctt tagccggaag tgctggcatt ttgtccaatt gagactcgtg caactggtca
     1081 gcgaactggt cgtagaaatc agccagtaca tcacaagact catatgtgtc aaccatagtt
     1141 tcgcgcactg ctttgaacag gttcgcagcg tcagccggaa tggtaccgaa ggagtcgtga
     1201 atcagtgcaa aagattcgat tccgtacttc tcgtgtgccc acactacagt cttacgaagg
     1261 tggctaccgt cttggctgtg tacaaagtta ggagcgatac cagactcctg tttgtgtgca
     1321 tcaatctcgc tatctttgtt ggtgttaatg gtaggctgta agcggaactg accgaggaac
     1381 atcaggttca agcgcgtctg aataggcttc ttgtattcct gccacacagg gaaaccatca
     1441 ggagttaccc aatgcacagc gcaacgcttg cgaagaatct ctccagtctt cttatctttg
     1501 acctcagcag ccagcagctt agcagcagac ttaagccagt tcattgcttc aaccgcagct
     1561 accaccgtca cgctcacaga ttcccaaatc agcttagcca tgtatccagc agcctgattc
     1621 ggctgagtga acatcagacc cttgccggaa tcaatagctg gctgaatggt atcttccagc
     1681 acttgttgac ggaagccgaa ctctttggac ccgtaagcca gcgtcatgac tgaacgctta
     1741 gtcacactgc gagtaacacc gtaagccagc cattgaccag ccagtgcctt agtgcccagc
     1801 ttgactttct cagagatttc accagtgttc tcatcggtca cggtaactac ttcgttatcg
     1861 gtcccattga ttgcgtctgc ttgtagaatc tcgttgactt tcttagcaac aatcccgtag
     1921 atgtcctgaa cggtttcact aggaagcaag ttaaccgcgc gaccacctac ctcatctcgg
     1981 agcatcgcgg agaagtgctg gatgccagag caagacccgt caaacgccag cggaagggag
     2041 cagttatagc tcaggccgtg gtgctgtacc ccagcgtact caaagcagaa cgcaaggaag
     2101 cagaacggag aatcttgctc agcccaccaa gtgttctcca gtggagactt agcgcaagcc
     2161 atgatgttct cgtggttttc ctcaatgaac ttgatgcgct cagggaacgg aaccttatcg
     2221 acacccgcac agtttgcacc gtggattttc agccagtagt aaccttcctt accgattggt
     2281 ttacctttcg ccagcgtaag cagtcctttg gtcatatcgt taccttgcgg gttgaacatt
     2341 gacacagcgt aaacacgacc gcgccagtcc atgttgtaag ggaaccagat ggccttatgg
     2401 ttagcaaact tattggcttg ctcaagcatg aactcaaggc tgatacggcg agacttgcga
     2461 gccttgtcct tgcggtacac agcagcggca gcacgtttcc acgcggtgag agcctcagga
     2521 ttcatgtcga tgtcttccgg tttcatcggg agttcttcac gctcaatcgc agggatgtcc
     2581 tcgaccggac aatgcttcca cttggtgatt acgttggcga ccgctaggac tttcttgttg
     2641 attttccatg cggtgttttg cgcaatgtta atcgctttgt acacctcagg catgtaaacg
     2701 tcttcgtagc gcatcagtgc tttcttactg tgagtacgca ccagcgccag aggacgacga
     2761 ccgttagccc aatagccacc accagtaatg ccagtccacg gcttaggagg aactacgcaa
     2821 ggttggaaca tcggagagat gccagccagc gcacctgcac gggttgcgat agcctcagcg
     2881 tattcaggtg cgagttcgat agtctcagag tcttgaccta ctacgccagc attttggcgg
     2941 tgtaagctaa ccattccggt tgactcaatg agcatctcga tgcagcgtac tcctacatga
     3001 atagagtctt ccttatgcca cgaagaccac gcctcgccac cgagtagacc cttagagagc
     3061 atgtcagcct cgacaacttg cataaatgct ttcttgtaga cgtgccctac gcgcttgttg
     3121 agttgttcct caacgttttt cttgaagtgc ttagcttcaa ggtcacggat acgaccgaag
     3181 cgagcctcgt cctcaatggc ccgaccgatt gcgcttgcta cagcctgaac ggttgtattg
     3241 tcagcactgg ttaggcaagc cagagtggtc ttaatggtga tgtacgctac ggcttccggc
     3301 ttgatttctt gcaggaactg gaaggctgtc gggcgcttgc cgcgcttagc tttcacttcc
     3361 tcaaaccagt cgttgatgcg tgcaatcatc ttagggagta gggtagtgat gagaggcttg
     3421 gcggcagcgt tatccgcaac ctcaccagct ttaagttgac gctcaaacat cttgcggaag
     3481 cgtgcttcac ccatctcgta agactcatgc tcaagggcca actgttcgcg agctaaacgc
     3541 tcaccgtaat ggtcagccag agtgttgaac gggatagcag ccagttcgat gtcagagaag
     3601 tcgttcttag cgatgttaat cgtgttcatt tagtgcctct tccagttagt aaatccggga
     3661 tcagatcccg aacgccatca aaaataattc gcgtctggcc ttcctgtagc cagctttcat
     3721 caacattaaa tgtgagcgag taacaacccg tcggattctc cgtgggaaca aacggcggat
     3781 tgaccgtaat gggataggtt acgttggtgt agatgggcgc atcgtaaccg tgcatctgcc
     3841 agtttgaggg gacgacgaca gtatcggcct caggaagatc gcactccagc cagctttccg
     3901 gcaccgcttc tggtgccgga aaccaggcaa agcgccattc gccattcagg ctgcgcaact
     3961 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     4021 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     4081 cgacggccag tgaatccgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4141 ctcacaattc cacacattat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4201 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4261 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4321 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     4381 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     4441 atcctgtttg atggtggttg acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     4501 tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     4561 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     4621 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     4681 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     4741 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     4801 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     4861 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     4921 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     4981 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     5041 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     5101 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     5161 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     5221 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     5281 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     5341 ttttttgcac cattgtggat cctctagagt cgacgaatta gcttcaaaag cgctctgaag
     5401 ttcctatact ttctagagaa taggaacttc ggaataggaa cttcaagatc ccctgattcc
     5461 ctttgtcaac agcaatggat cgaattgaca taagcctgtt cggttcgtaa actgtaatgc
     5521 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
     5581 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttgtacagtc tatgcctcgg
     5641 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
     5701 gatgttacgc agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttagg
     5761 tggctcaagt atgggcatca ttcgcacatg taggctcggc cctgaccaag tcaaatccat
     5821 gcgggctgct cttgatcttt tcggtcgtga gttcggagac gtagccacct actcccaaca
     5881 tcagccggac tccgattacc tcgggaactt gctccgtagt aagacattca tcgcgcttgc
     5941 tgccttcgac caagaagcgg ttgttggcgc tctcgcggct tacgttctgc ccaggtttga
     6001 gcagccgcgt agtgagatct atatctatga tctcgcagtc tccggcgagc accggaggca
     6061 gggcattgcc accgcgctca tcaatctcct caagcatgag gccaacgcgc ttggtgctta
     6121 tgtgatctac gtgcaagcag attacggtga cgatcccgca gtggctctct atacaaagtt
     6181 gggcatacgg gaagaagtga tgcactttga tatcgaccca agtaccgcca cctaacaatt
     6241 cgttcaagcc gagatcggct tcccggccgc ggagttgttc ggtaaattgt cacaacgccg
     6301 cggccaattc gatctagaat tccattgagt aagtttttaa gcacatcagc ttcaaaagcg
     6361 ctctgaagtt cctatacttt ctagagaata ggaacttcgg aataggaact tcaagatccc
     6421 caattcgtcg acctgcaggc atgcaaggag atggcgccca acagtccccc ggccacgggg
     6481 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     6541 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     6601 ccggccacga tgcgtccggc gtagaggatc cgggtcccct ttgatagatt aaaaaggaaa
     6661 ggaggaaaga aataatggct cgtgtacagt ttaaacaacg tgaatctact gacgcaatct
     6721 ttgttcactg ctcggctacc aagccaagtc agaatgttgg tgtccgtgag attcgccagt
     6781 ggcacaaaga gcagggttgg ctcgatgtgg gataccactt tatcatcaag cgagacggta
     6841 ctgtggaggc aggacgagat gagatggctg taggctctca cgctaagggt tacaaccaca
     6901 actctatcgg cgtctgcctt gttggtggta tcgacgataa aggtaagttc gacgctaact
     6961 ttacgccagc ccaaatgcaa tcccttcgct cactgcttgt cacactgctg gctaagtacg
     7021 aaggcgctgg tcttcgcgcc catcatgagg tggcgccgaa ggcttgccct tcgttcgacc
     7081 ttaagcgttg gtgggagaag aacgaactgg tcacttctga ccgtggataa ttaattgaac
     7141 tcactaaagg gagaccacgc atg
//
