LOCUS       V002399                 8856 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002399
VERSION     V002399
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8856)
  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 8856)
  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 8856)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8856)
  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..8856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(437..889)
                     /label="T7 lysozyme"
                     /note="lysozyme from bacteriophage T7"
     protein_bind    1152..1199
                     /label="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1332..1862)
                     /label="GmR"
                     /note="gentamycin acetyltransferase"
     promoter        complement(2051..2079)
                     /label="Pc promoter"
                     /note="class 1 integron promoter"
     misc_feature    2120..2167
                     /note="FRT; Flp-recombination-target"
     protein_bind    2120..2167
                     /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)."
     CDS             2248..3327
                     /label="lacI"
                     /note="lac repressor"
     protein_bind    3343..3364
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3379..3409
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    3417..3433
                     /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)."
     CDS             3453..3626
                     /label="lacZ-alpha"
                     /note="LacZ-alpha fragment of beta-galactosidase"
     CDS             3935..6583
                     /note="T7 RNA polymerase from Escherichia phage T7.
                     Accession#: P00573"
                     /label="T7 RNA polymerase"
     primer_bind     complement(6635..6651)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6659..6675)
                     /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(6683..6713)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6728..6749)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7037..7625)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7799..8656)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8657..8761)
                     /label="AmpR promoter"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccctttag
      421 tgagttcaat taattatcca cggtcagaag tgaccagttc gttcttctcc caccaacgct
      481 taaggtcgaa cgaagggcaa gccttcggcg ccacctcatg atgggcgcga agaccagcgc
      541 cttcgtactt agccagcagt gtgacaagca gtgagcgaag ggattgcatt tgggctggcg
      601 taaagttagc gtcgaactta cctttatcgt cgataccacc aacaaggcag acgccgatag
      661 agttgtggtt gtaaccctta gcgtgagagc ctacagccat ctcatctcgt cctgcctcca
      721 cagtaccgtc tcgcttgatg ataaagtggt atcccacatc gagccaaccc tgctctttgt
      781 gccactggcg aatctcacgg acaccaacat tctgacttgg cttggtagcc gagcagtgaa
      841 caaagattgc gtcagtagat tcacgttgtt taaactgtac acgagccatt atttctttcc
      901 tcctttcctt tttaatctat caaaggggac ccggatcctc tacgccggac gcatcgtggc
      961 cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg
     1021 ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc
     1081 aggccccgtg gccgggggac tgttgggcgc catctccttg catgcctgca ggtcgacgaa
     1141 ttggggatct tgaagttcct attccgaagt tcctattctc tagaaagtat aggaacttca
     1201 gagcgctttt gaagctgatg tgcttaaaaa cttactcaat ggaattctag atcgaattgg
     1261 ccgcggcgtt gtgacaattt accgaacaac tccgcggccg ggaagccgat ctcggcttga
     1321 acgaattgtt aggtggcggt acttgggtcg atatcaaagt gcatcacttc ttcccgtatg
     1381 cccaactttg tatagagagc cactgcggga tcgtcaccgt aatctgcttg cacgtagatc
     1441 acataagcac caagcgcgtt ggcctcatgc ttgaggagat tgatgagcgc ggtggcaatg
     1501 ccctgcctcc ggtgctcgcc ggagactgcg agatcataga tatagatctc actacgcggc
     1561 tgctcaaacc tgggcagaac gtaagccgcg agagcgccaa caaccgcttc ttggtcgaag
     1621 gcagcaagcg cgatgaatgt cttactacgg agcaagttcc cgaggtaatc ggagtccggc
     1681 tgatgttggg agtaggtggc tacgtctccg aactcacgac cgaaaagatc aagagcagcc
     1741 cgcatggatt tgacttggtc agggccgagc ctacatgtgc gaatgatgcc catacttgag
     1801 ccacctaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct gctgcgtaac
     1861 atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt gctgcttgga
     1921 tgcccgaggc atagactgta caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc
     1981 gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta
     2041 cttgcattac agtttacgaa ccgaacaggc ttatgtcaat tcgatccatt gctgttgaca
     2101 aagggaatca ggggatcttg aagttcctat tccgaagttc ctattctcta gaaagtatag
     2161 gaacttcaga gcgcttttga agctaattcg tcgactctag aggatccaca atggtgcaaa
     2221 aaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     2281 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     2341 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     2401 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     2461 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     2521 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
     2581 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     2641 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     2701 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
     2761 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
     2821 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     2881 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     2941 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     3001 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     3061 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgtcaaccac catcaaacag
     3121 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     3181 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     3241 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     3301 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     3361 tcattaggca ccccaggctt tacactttat gcttccggct cgtataatgt gtggaattgt
     3421 gagcggataa caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg
     3481 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     3541 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3601 aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg
     3661 tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa
     3721 actggcagat gcacggttac gatgcgccca tctacaccaa cgtaacctat cccattacgg
     3781 tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg
     3841 ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc gttcgggatc
     3901 tgatcccgga tttactaact ggaagaggca ctaaatgaac acgattaaca tcgctaagaa
     3961 cgacttctct gacatcgaac tggctgctat cccgttcaac actctggctg accattacgg
     4021 tgagcgttta gctcgcgaac agttggccct tgagcatgag tcttacgaga tgggtgaagc
     4081 acgcttccgc aagatgtttg agcgtcaact taaagctggt gaggttgcgg ataacgctgc
     4141 cgccaagcct ctcatcacta ccctactccc taagatgatt gcacgcatca acgactggtt
     4201 tgaggaagtg aaagctaagc gcggcaagcg cccgacagcc ttccagttcc tgcaagaaat
     4261 caagccggaa gccgtagcgt acatcaccat taagaccact ctggcttgcc taaccagtgc
     4321 tgacaataca accgttcagg ctgtagcaag cgcaatcggt cgggccattg aggacgaggc
     4381 tcgcttcggt cgtatccgtg accttgaagc taagcacttc aagaaaaacg ttgaggaaca
     4441 actcaacaag cgcgtagggc acgtctacaa gaaagcattt atgcaagttg tcgaggctga
     4501 catgctctct aagggtctac tcggtggcga ggcgtggtct tcgtggcata aggaagactc
     4561 tattcatgta ggagtacgct gcatcgagat gctcattgag tcaaccggaa tggttagctt
     4621 acaccgccaa aatgctggcg tagtaggtca agactctgag actatcgaac tcgcacctga
     4681 atacgctgag gctatcgcaa cccgtgcagg tgcgctggct ggcatctctc cgatgttcca
     4741 accttgcgta gttcctccta agccgtggac tggcattact ggtggtggct attgggctaa
     4801 cggtcgtcgt cctctggcgc tggtgcgtac tcacagtaag aaagcactga tgcgctacga
     4861 agacgtttac atgcctgagg tgtacaaagc gattaacatt gcgcaaaaca ccgcatggaa
     4921 aatcaacaag aaagtcctag cggtcgccaa cgtaatcacc aagtggaagc attgtccggt
     4981 cgaggacatc cctgcgattg agcgtgaaga actcccgatg aaaccggaag acatcgacat
     5041 gaatcctgag gctctcaccg cgtggaaacg tgctgccgct gctgtgtacc gcaaggacaa
     5101 ggctcgcaag tctcgccgta tcagccttga gttcatgctt gagcaagcca ataagtttgc
     5161 taaccataag gccatctggt tcccttacaa catggactgg cgcggtcgtg tttacgctgt
     5221 gtcaatgttc aacccgcaag gtaacgatat gaccaaagga ctgcttacgc tggcgaaagg
     5281 taaaccaatc ggtaaggaag gttactactg gctgaaaatc cacggtgcaa actgtgcggg
     5341 tgtcgataag gttccgttcc ctgagcgcat caagttcatt gaggaaaacc acgagaacat
     5401 catggcttgc gctaagtctc cactggagaa cacttggtgg gctgagcaag attctccgtt
     5461 ctgcttcctt gcgttctgct ttgagtacgc tggggtacag caccacggcc tgagctataa
     5521 ctgctccctt ccgctggcgt ttgacgggtc ttgctctggc atccagcact tctccgcgat
     5581 gctccgagat gaggtaggtg gtcgcgcggt taacttgctt cctagtgaaa ccgttcagga
     5641 catctacggg attgttgcta agaaagtcaa cgagattcta caagcagacg caatcaatgg
     5701 gaccgataac gaagtagtta ccgtgaccga tgagaacact ggtgaaatct ctgagaaagt
     5761 caagctgggc actaaggcac tggctggtca atggctggct tacggtgtta ctcgcagtgt
     5821 gactaagcgt tcagtcatga cgctggctta cgggtccaaa gagttcggct tccgtcaaca
     5881 agtgctggaa gataccattc agccagctat tgattccggc aagggtctga tgttcactca
     5941 gccgaatcag gctgctggat acatggctaa gctgatttgg gaatctgtga gcgtgacggt
     6001 ggtagctgcg gttgaagcaa tgaactggct taagtctgct gctaagctgc tggctgctga
     6061 ggtcaaagat aagaagactg gagagattct tcgcaagcgt tgcgctgtgc attgggtaac
     6121 tcctgatggt ttccctgtgt ggcaggaata caagaagcct attcagacgc gcttgaacct
     6181 gatgttcctc ggtcagttcc gcttacagcc taccattaac accaacaaag atagcgagat
     6241 tgatgcacac aaacaggagt ctggtatcgc tcctaacttt gtacacagcc aagacggtag
     6301 ccaccttcgt aagactgtag tgtgggcaca cgagaagtac ggaatcgaat cttttgcact
     6361 gattcacgac tccttcggta ccattccggc tgacgctgcg aacctgttca aagcagtgcg
     6421 cgaaactatg gttgacacat atgagtcttg tgatgtactg gctgatttct acgaccagtt
     6481 cgctgaccag ttgcacgagt ctcaattgga caaaatgcca gcacttccgg ctaaaggtaa
     6541 cttgaacctc cgtgacatct tagagtcgga cttcgcgttc gcgtaaggat cctctagagt
     6601 cgacctgcag gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6661 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6721 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6781 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6841 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6901 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6961 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7021 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7081 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7141 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7201 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7261 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7321 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7381 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7441 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     7501 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7561 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7621 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7681 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7741 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7801 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7861 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7921 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7981 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     8041 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8101 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8161 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8221 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8281 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8341 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8401 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8461 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8521 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8581 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8641 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8701 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8761 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     8821 cctataaaaa taggcgtatc acgaggccct ttcgtc
//
