LOCUS       40924_43418        8193 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTip-LNH2 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8193)
  AUTHORS   Nakashima N, Tamura T.
  TITLE     A novel system for expressing recombinant proteins over a wide 
            temperature range from 4 to 35 degrees C
  JOURNAL   Biotechnol. Bioeng. 86 (2), 136-148 (2004)
  PUBMED    15052633
REFERENCE   2  (bases 1 to 8193)
  AUTHORS   Nakashima N, Tamura T.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-OCT-2002) Nobutaka Nakashima, National Institute of 
            Advanced Industrial Science and Technology; Tsukisamu-Higashi 
            2-17-2-1, Sapporo, Toyohira-ku 062-8517, Japan 
            (E-mail:n-nakashima@aist.go.jp, Tel:81-11-857-8937)
REFERENCE   3  (bases 1 to 8193)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8193)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol.
            Bioeng."; date: "2004"; volume: "86"; issue: "2"; pages: "136-148"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-OCT-2002) Nobutaka Nakashima, National Institute of Advanced 
            Industrial Science and Technology"; volume: " Tsukisamu-Higashi 
            2-17-2-1, Sapporo, Toyohira-ku 062-8517, Japan 
            (E-mail:n-nakashima@aist.go.jp, Tel"; pages: "81-11-857-8937"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             170..976
                     /gene="tsnR"
                     /label=tsnR
                     /note="23S rRNA (adenosine(1067)-2'-O)-methyltransferase
                     from Streptomyces azureus. Accession#: P18644"
     regulatory      1145..1324
                     /gene="Tuf1"
                     /regulatory_class="promoter"
     gene            1145..1324
                     /gene="Tuf1"
                     /label=Tuf1
     CDS             1325..2512
                     /label=TcR
                     /note="tetracycline efflux protein"
     regulatory      2728..2903
                     /gene="ThcA"
                     /regulatory_class="promoter"
     gene            2728..2903
                     /gene="ThcA"
                     /label=ThcA
     CDS             2904..3662
                     /gene="tipA"
                     /label=tipA
                     /note="HTH-type transcriptional activator TipA from
                     Streptomyces lividans. Accession#: P0A4T9"
     promoter        3843..3947
                     /label=AmpR promoter
     CDS             3948..4805
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4979..5567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      complement(5818..5998)
                     /gene="ThcA"
                     /regulatory_class="terminator"
     gene            complement(5818..5998)
                     /gene="ThcA"
                     /label=ThcA
     CDS             complement(6063..6080)
                     /label=6xHis
                     /note="6xHis affinity tag"
     RBS             complement(6094..6116)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      complement(6130..6253)
                     /gene="TipA"
                     /regulatory_class="promoter"
     CDS             6792..7679
                     /codon_start=1
                     /gene="RepA"
                     /label=RepA
                     /protein_id="BAD11248.1"
                     /translation="MKSATGGVLSDRQWADAVLLPHRPFATNNLQRGQYRMSRDDALAM
                     RYVEHSPHALLGSIVIDCDHVDAAMRAFEQPSDHPAPNWVAQSPSGRAHIGWWLGPNHV
                     CRTDSARLTPLRYAHRIETGLKISVGGDFAYGGQLTKNPIHPDWETIYGPATPYTLRQL
                     ATIHTPRQMPRRPDRAVGLGRNVTMFDATRRWAYPQWWQHRNGTGRDWDHLVLQHCHAV
                     NTEFTTPLPFTEVRATAQSISKWIWRNFTEEQYRARQAHLGQKGGKATTLAKQEAVRNN
                     ARKYDEHTMREAII"
     gene            6792..7679
                     /gene="RepA"
                     /label=RepA
     CDS             7679..7963
                     /codon_start=1
                     /gene="RepB"
                     /label=RepB
                     /protein_id="BAD11249.1"
                     /translation="MGGAKNPVRRKMTAAAAAEKFGASTRTIQRLFAEPRDDYLGRAKA
                     RRDKAVELRKQGLKYREIAEAMELSTGIVGRLLHDARRHGEISAEDLSA"
     gene            7679..7963
                     /gene="RepB"
                     /label=RepB
ORIGIN
        1 gagctcgacc gcgcgggtcc cggacgggga agagcgggga gctttgccag agagcgacga
       61 cttccccttg cgttggtgat tgccggtcag ggcagccatc cgccatcgtc gcgtagggtg
      121 tcacacccca ggaatcgcgt cactgaacac agcagccggt aggacgacca tgactgagtt
      181 ggacaccatc gcaaatccgt ccgatcccgc ggtgcagcgg atcatcgatg tcaccaagcc
      241 gtcacgatcc aacataaaga caacgttgat cgaggacgtc gagcccctca tgcacagcat
      301 cgcggccggg gtggagttca tcgaggtcta cggcagcgac agcagtcctt ttccatctga
      361 gttgctggat ctgtgcgggc ggcagaacat accggtccgc ctcatcgact cctcgatcgt
      421 caaccagttg ttcaaggggg agcggaaggc caagacattc ggcatcgccc gcgtccctcg
      481 cccggccagg ttcggcgata tcgcgagccg gcgtggggac gtcgtcgttc tcgacggggt
      541 gaagatcgtc gggaacatcg gcgcgatagt acgcacgtcg ctcgcgctcg gagcgtcggg
      601 gatcatcctg gtggacagtg acatcaccag catcgcggac cggcgtctcc aaagggccag
      661 ccgaggttac gtcttctccc ttcccgtcgt tctctccggt cgcgaggagg ccatcgcctt
      721 cattcgggac agcggtatgc agctgatgac gctcaaggcg gatggcgaca tttccgtgaa
      781 ggaactcggg gacaatccgg atcggctggc cttgctgttc ggcagcgaaa agggtgggcc
      841 ttccgacctg ttcgaggagg cgtcttccgc ctcggtttcc atccccatga tgagccagac
      901 cgagtctctc aacgtttccg tttccctcgg aatcgcgctg cacgagagga tcgacaggaa
      961 tctcgcggcc aaccgataag cgcctctgtt cctcggacgc tcggttcctc gacctcgatt
     1021 cgtcagtgat gatcacctca cacggcagcg atcaccactg acatatcgag gtcaacggtc
     1081 gtggtccggg cgggcactcc tcgaaggcgc ggccgacgcc cttgaacgac tcgatgactc
     1141 tagagtaacg ggctactccg tttaacggac cccgttctca cgctttaggc ttgaccccgg
     1201 agcctgcatg gggcattccg ccgtgaaccc ggtggaatgc ccccggcacc cgggctttcc
     1261 agcaaagatc acctggcgcc gatgagtaag gcgtacagaa ccactccaca ggaggaccgt
     1321 cgagatgaaa tctaacaatg cgctcatcgt catcctcggc accgtcaccc tggatgctgt
     1381 aggcataggc ttggttatgc cggtactgcc gggcctcttg cgggatatcg tccattccga
     1441 cagcatcgcc agtcactatg gcgtgctgct agcgctatat gcgttgatgc aatttctatg
     1501 cgcacccgtt ctcggagcac tgtccgaccg ctttggccgc cgcccagtcc tgctcgcttc
     1561 gctacttgga gccactatcg actacgcgat catggcgacc acacccgtcc tgtggattct
     1621 ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta
     1681 tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg
     1741 tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt
     1801 gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt
     1861 cctaatgcag gagtcgcata agggagagcg tcgtccgatg cccttgagag ccttcaaccc
     1921 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     1981 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga
     2041 ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca
     2101 cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc
     2161 cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg
     2221 aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc
     2281 gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc
     2341 gctcgcggct cttaccagcc taacttcgat cattggaccg ctgatcgtca cggcgattta
     2401 tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct
     2461 tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga
     2521 agccggcggc acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg
     2581 cggagaactg tgaatgcgca aaccaaccct tggcagaaca tatccatcgc gtccgccatc
     2641 tccagcagcc gcacgcggcg catctcgggc agcgttgggt cctggccacg ggtgcgcatg
     2701 atcgtgctcc tgtcgttgag gactagaatt gatctcctcg accgccaatt gggcatctga
     2761 gaatcatctg cgtttctcgc acgcaacgta cttgcaacgt tgcaactcct agtgttgtga
     2821 atcacacccc accggggggt gggattgcag tcaccgattt ggtgggtgcg cccaggaaga
     2881 tcacgtttac ataggagctt gcaatgagct actccgtggg acaggtggcc ggcttcgccg
     2941 gagtgacggt gcgcacgctg caccactacg acgacatcgg cctgctcgta ccgagcgagc
     3001 gcagccacgc gggccaccgg cgctacagcg acgccgacct cgaccggctg cagcagatcc
     3061 tgttctaccg ggagctgggc ttcccgctcg acgaggtcgc cgccctgctc gacgacccgg
     3121 ccgcggaccc gcgcgcgcac ctgcgccgcc agcacgagct gctgtccgcc cggatcggga
     3181 aactgcagaa gatggcggcg gccgtggagc aggcgatgga ggcacgcagc atgggaatca
     3241 acctcacccc ggaggagaag ttcgaggtct tcggcgactt cgaccccgac cagtacgagg
     3301 aggaggtccg ggaacgctgg gggaacaccg acgcctaccg ccagtccaag gagaagaccg
     3361 cctcgtacac caaggaggac tggcagcgca tccaggacga ggccgacgag ctcacccggc
     3421 gcttcgtcgc cctgatggac gcgggtgagc ccgccgactc cgagggggcg atggacgccg
     3481 ccgaggacca ccggcagggc atcgcccgca accactacga ctgcgggtac gagatgcaca
     3541 cctgcctggg cgagatgtac gtgtccgacg aacgtttcac gcgaaacatc gacgccgcca
     3601 agccgggcct cgccgcctac atgcgcgacg cgatcctcgc caacgccgtc cggcacaccc
     3661 cctgagcggt ggtcgtggcc cgggtctccc gcccggtctc accccacggc tcactcccgg
     3721 gccacgacca ccgccgtccc gtacgcgcac acctcggtgc ccacgtccgc cgcctccgtc
     3781 acgtcgaaac ggaagatccc cgggtaccga gctcgtcagg tggcactttt cggggaaatg
     3841 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3901 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3961 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4021 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4081 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4141 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4201 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4261 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4321 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4381 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4441 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4501 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4561 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4621 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4681 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4741 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4801 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4861 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4921 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4981 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5041 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5101 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5161 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5221 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5281 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5341 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5401 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5461 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5521 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5581 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5641 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5701 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     5761 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gactagagtc
     5821 ccgctgaggc ggcgtagcag gtcagccgcc ccagcggtgg tcaccaaccg gggtggaacg
     5881 gcgccggtat cgggtgtgtc cgtggcgctc attccaacct ccgtgtgttt gtgcaggttt
     5941 cgcgtgttgc agtccctcgc accggcaccc gcagcgaggg gctcacgggt gccggtgggt
     6001 cgactagttc atcctcgaga tctaagcttg gatccgcggc cgctacgtag aattcccatg
     6061 gcgtgatggt gatggtgatg gcccatatgt atatctcctt cttaaagtta aacaaaatta
     6121 tttctagacg ccgtccacgc tgcctcctca cgtgacgtga ggtgcaagcc cggacgttcc
     6181 gcgtgccacg ccgtgagccg ccgcgtgccg tcggctccct cagcccgggc ggccgtggga
     6241 gcccgcctcg atatgtacac ccgagaagct cccagcgtcc tcctgggccg cgatactcga
     6301 ccaccacgca cgcacaccgc actaacgatt cggccggcgc tcgattcggc cggcgctcga
     6361 ttcggccggc gctcgattcg gccggcgctc gattcggccg gcgctcgatt cggccgagca
     6421 gaagagtgaa caaccaccga ccacgcttcc gctctgcgcg ccgtacccga cctacctccc
     6481 gcagctcgaa gcagctcccg ggagtaccgc cgtactcacc cgcctgtgct caccatccac
     6541 cgacgcaaag cccaacccga gcacacctct tgcaccaagg tgccgaccgt ggctttccgc
     6601 tcgcagggtt ccagaagaaa tcgaacgatc cagcgcggca aggttcaaaa agcaggggtt
     6661 ggtggggagg aggttttggg gggtgtcgcc gggatacctg atatggcttt gttttgcgta
     6721 gtcgaataat tttccatata gcctcggcgc gtcggactcg aatagttgat gtgggcgggc
     6781 acagttgccc catgaaatcc gcaacggggg gcgtgctgag cgatcggcaa tgggcggatg
     6841 cggtgttgct tccgcaccgg ccgttcgcga cgaacaacct ccaacgaggt cagtaccgga
     6901 tgagccgcga cgacgcattg gcaatgcggt acgtcgagca ttcaccgcac gcgttgctcg
     6961 gatctatcgt catcgactgc gatcacgttg acgccgcgat gcgcgcattc gagcaaccat
     7021 ccgaccatcc ggcgccgaac tgggtcgcac aatcgccgtc cggccgcgca cacatcggat
     7081 ggtggctcgg ccccaaccac gtgtgccgca ccgacagcgc ccgactgacg ccactgcgct
     7141 acgcccaccg catcgaaacc ggcctcaaga tcagcgtcgg cggcgatttc gcgtatggcg
     7201 ggcaactgac caaaaacccg attcaccccg attgggagac gatctacggc ccggccaccc
     7261 cgtacacatt gcggcagctg gccaccatcc acacaccccg gcagatgccg cgtcggcccg
     7321 atcgggccgt gggcctgggc cgcaacgtca ccatgttcga cgccacccgg cgatgggcat
     7381 acccgcagtg gtggcaacac cgaaacggaa ccggccgcga ctgggaccat ctcgtcctgc
     7441 agcactgcca cgccgtcaac accgagttca cgacaccact gccgttcacc gaagtacgcg
     7501 ccaccgcgca atccatctcc aaatggatct ggcgcaattt caccgaagaa cagtaccgag
     7561 cccgacaagc gcatctcggt caaaaaggcg gcaaggcaac gacactcgcc aaacaagaag
     7621 ccgtccgaaa caatgcaaga aagtacgacg aacatacgat gcgagaggcg attatctgat
     7681 gggcggagcc aaaaatccgg tgcgccgaaa gatgacggca gcagcagcag ccgaaaaatt
     7741 cggtgcctcc actcgcacaa tccaacgctt gtttgctgag ccgcgtgacg attacctcgg
     7801 ccgtgcgaaa gctcgccgtg acaaagctgt cgagctgcgg aagcaggggt tgaagtaccg
     7861 ggaaatcgcc gaagcgatgg aactctcgac cgggatcgtc ggccgattac tgcacgacgc
     7921 ccgcaggcac ggcgagattt cagcggagga tctgtcggcg taaccaagtc agcgggttgt
     7981 cgggttccgg ccggcgctcg gcactcggac cggccggcgg atggtgttct gcctctggcg
     8041 cagcgtcagc taccgccgaa ggcctgtcat cgaccggctt cgactgaagt atgagcaacg
     8101 tcacagcctg tgattggatg atccgctcac gctcgaccgc tacctgttca gctgccgccc
     8161 gctgggcatg agcaacggcc aactctcgtt caa
//
