LOCUS       40924_43403        8361 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTip-LCH2.2 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8361)
  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 8361)
  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 8361)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8361)
  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..8361
                     /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"
     CDS             1541..2716
                     /codon_start=1
                     /gene="Chlr"
                     /product="chloramphenicol resistance protein"
                     /label=Chlr
                     /protein_id="BAD11299.1"
                     /translation="VPFAIYVLGLAVFAQGTSEFMLSGLIPDMARDLGVSVPAAGLLTS
                     AFAVGMIIGAPLMAIASMRWPRRRALLTFLITFMLVHVIGALTSSFEVLLVTRIVGALA
                     NAGFLAVALGAAMAMVPADMKGRATSVLLGGVTIACVAGVPGGAFLGEMWGWRAAFWAV
                     VVISAPAVVAIMFATPAEPLAESTPNAKRELSSLRSRKLQLMLVLGALINGATFCSFTY
                     MAPTLTDISGFDSRWIPLLLGLFGLGSFIGVSVGGRLADTRPFQLLAVGSAALLTGWIV
                     FALTASHPAVTLVMLFVQGALSFAVGSTLISQVLYAADAAPTLGGSFATAAFNVGAALG
                     PALGGLAIGMGLSYRAPLWTSAALVTLAIVIGAATLSLWRRPASVHESVPA"
     gene            1541..2716
                     /gene="Chlr"
                     /label=Chlr
     regulatory      2905..3080
                     /gene="ThcA"
                     /regulatory_class="promoter"
     gene            2905..3080
                     /gene="ThcA"
                     /label=ThcA
     CDS             3081..3839
                     /gene="tipA"
                     /label=tipA
                     /note="HTH-type transcriptional activator TipA from
                     Streptomyces lividans. Accession#: P0A4T9"
     promoter        4020..4124
                     /label=AmpR promoter
     CDS             4125..4982
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5156..5744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      complement(5995..6175)
                     /gene="ThcA"
                     /regulatory_class="terminator"
     gene            complement(5995..6175)
                     /gene="ThcA"
                     /label=ThcA
     CDS             complement(6189..6206)
                     /label=6xHis
                     /note="6xHis affinity tag"
     RBS             complement(6262..6284)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      complement(6298..6421)
                     /gene="TipA"
                     /regulatory_class="promoter"
     CDS             6960..7847
                     /codon_start=1
                     /gene="RepA"
                     /label=RepA
                     /protein_id="BAD11302.1"
                     /translation="MKSATGGVLSDRQWADAVLLPHRPFATNNLQRGQYRMSRDDALAM
                     RYVEHSPHALLGSIVIDCDHVDAAMRAFEQPSDHPAPNWVAQSPSGRAHIGWWLGPNHV
                     CRTDSARLTPLRYAHRIETGLKISVGGDFAYGGQLTKNPIHPDWETIYGPATPYTLRQL
                     ATIHTPRQMPRRPDRAVGLGRNVTMFDATRRWAYPQWWQHRNGTGRDWDHLVLQHCHAV
                     NTEFTTPLPFTEVRATAQSISKWIWRNFTEEQYRARQAHLGQKGGKATTLAKQEAVRNN
                     ARKYDEHTMREAII"
     gene            6960..7847
                     /gene="RepA"
                     /label=RepA
     CDS             7847..8131
                     /codon_start=1
                     /gene="RepB"
                     /label=RepB
                     /protein_id="BAD11303.1"
                     /translation="MGGAKNPVRRKMTAAAAAEKFGASTRTIQRLFAEPRDDYLGRAKA
                     RRDKAVELRKQGLKYREIAEAMELSTGIVGRLLHDARRHGEISAEDLSA"
     gene            7847..8131
                     /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 tagacgcatc cgaaacctcc accccactca cctagtccga catccgtacc ttggaaaccg
     1201 acctgtattg gcatttcagt tggacatcga ccagtggcgt tgctaggttc aagaccatgt
     1261 ccagcccgaa ggcgtccaga ctctagccac cggaggtagt ccggtggcca catcccgtcg
     1321 cgcccgaacg tcacgctctt gtgtggcctt cccttgttgt ttgcgatcag tggcacacct
     1381 ctaccgtctg aatttcgagt ctggcctcgg ctgcgcacat ctcgcactgt gacgctgtca
     1441 ggtcacccgc ttcgcggcta ccagttcctt tcatcgaatc gagcttccgg tgccgccgcg
     1501 cagcctccct gaccatcctc agattttatg gagtctcgca gtgcctttcg ctatctacgt
     1561 cctcgggctt gctgtcttcg cccagggcac atccgagttc atgttgtccg gactcatacc
     1621 ggacatggcc cgtgacctcg gggtttcggt ccccgccgcc ggactcctca cctccgcctt
     1681 cgcggtcggg atgatcatcg gcgctccgtt gatggctatc gccagcatgc ggtggccccg
     1741 gcgacgcgcc cttctgacat tcctcatcac gttcatgctg gtccacgtca tcggcgcgct
     1801 caccagcagc ttcgaggtct tgctggtcac acgcatcgtg ggagccctcg ccaatgccgg
     1861 attcttggca gtggccctgg gggcggcgat ggcgatggtg cccgccgaca tgaaagggcg
     1921 cgccacgtcc gtcctcctcg gcggtgtcac gatcgcatgt gtagccggtg ttcccggggg
     1981 cgccttcctg ggtgaaatgt ggggctggcg tgcagcgttc tgggctgtcg tcgtcatctc
     2041 cgcccctgca gtggtggcga ttatgttcgc caccccggcc gagccgcttg cagagtccac
     2101 accgaatgcc aagcgtgaac tgtcctcact gcgctcacgc aagctccagc tcatgcttgt
     2161 cctcggggcg ctgatcaacg gcgcaacgtt ctgttcgttc acgtacatgg cgcccacgct
     2221 caccgacatc tccggtttcg actcccgttg gattccgttg ctgctggggc tgttcgggct
     2281 cggatcgttc atcggtgtca gcgtcggagg caggctcgcc gacacccggc cgttccaact
     2341 gctcgctgtc gggtccgcag cactgttgac gggatggatc gtcttcgctc tcacggcatc
     2401 ccaccccgcg gtgacattgg tgatgctgtt cgtgcagggc gctttgtcct tcgcggtcgg
     2461 ctcgactttg atctcccagg tgctctacgc cgccgacgcg gcaccgacct tgggtggatc
     2521 gttcgcgacg gccgcgttca acgtcggtgc tgcactggga ccggcgctcg gcgggttggc
     2581 gatcggcatg ggtctgagct accgcgcccc gctctggacg agcgccgcgc tggtgacact
     2641 cgcgatcgtc atcggcgcag ccaccttgtc tctgtggcgg cgaccagcgt ctgtccacga
     2701 atctgtcccc gcctgaccag aaaccaggat ctgtgagtgt ggtgactgat ctgtgcacgc
     2761 tcagcagtca ccgcgcgctc gcgtcgtacc gagggccagc gccaacaggt gtgtggagct
     2821 ctgcccctgc ctctttcacg cgaactcact gttcagtgcg gcgatacgtg ctcggtgagt
     2881 tccactacag cgaccatgac tagaattgat ctcctcgacc gccaattggg catctgagaa
     2941 tcatctgcgt ttctcgcacg caacgtactt gcaacgttgc aactcctagt gttgtgaatc
     3001 acaccccacc ggggggtggg attgcagtca ccgatttggt gggtgcgccc aggaagatca
     3061 cgtttacata ggagcttgca atgagctact ccgtgggaca ggtggccggc ttcgccggag
     3121 tgacggtgcg cacgctgcac cactacgacg acatcggcct gctcgtaccg agcgagcgca
     3181 gccacgcggg ccaccggcgc tacagcgacg ccgacctcga ccggctgcag cagatcctgt
     3241 tctaccggga gctgggcttc ccgctcgacg aggtcgccgc cctgctcgac gacccggccg
     3301 cggacccgcg cgcgcacctg cgccgccagc acgagctgct gtccgcccgg atcgggaaac
     3361 tgcagaagat ggcggcggcc gtggagcagg cgatggaggc acgcagcatg ggaatcaacc
     3421 tcaccccgga ggagaagttc gaggtcttcg gcgacttcga ccccgaccag tacgaggagg
     3481 aggtccggga acgctggggg aacaccgacg cctaccgcca gtccaaggag aagaccgcct
     3541 cgtacaccaa ggaggactgg cagcgcatcc aggacgaggc cgacgagctc acccggcgct
     3601 tcgtcgccct gatggacgcg ggtgagcccg ccgactccga gggggcgatg gacgccgccg
     3661 aggaccaccg gcagggcatc gcccgcaacc actacgactg cgggtacgag atgcacacct
     3721 gcctgggcga gatgtacgtg tccgacgaac gtttcacgcg aaacatcgac gccgccaagc
     3781 cgggcctcgc cgcctacatg cgcgacgcga tcctcgccaa cgccgtccgg cacaccccct
     3841 gagcggtggt cgtggcccgg gtctcccgcc cggtctcacc ccacggctca ctcccgggcc
     3901 acgaccaccg ccgtcccgta cgcgcacacc tcggtgccca cgtccgccgc ctccgtcacg
     3961 tcgaaacgga agatccccgg gtaccgagct cgtcaggtgg cacttttcgg ggaaatgtgc
     4021 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4081 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4141 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4201 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4261 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4321 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4381 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4441 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4501 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4561 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4621 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     4681 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4741 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4801 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4861 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4921 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4981 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5041 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5101 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5161 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5221 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5281 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5341 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5401 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5461 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5521 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5581 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5641 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5701 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5761 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5821 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     5881 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     5941 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac tagagtcccg
     6001 ctgaggcggc gtagcaggtc agccgcccca gcggtggtca ccaaccgggg tggaacggcg
     6061 ccggtatcgg gtgtgtccgt ggcgctcatt ccaacctccg tgtgtttgtg caggtttcgc
     6121 gtgttgcagt ccctcgcacc ggcacccgca gcgaggggct cacgggtgcc ggtgggtcga
     6181 ctagttcagt gatggtgatg gtgatgtcct cgagatctaa gcttggatcc gcggccgcta
     6241 cgtagaattc ccatatgtat atctccttct taaagttaaa caaaattatt tctagacgcc
     6301 gtccacgctg cctcctcacg tgacgtgagg tgcaagcccg gacgttccgc gtgccacgcc
     6361 gtgagccgcc gcgtgccgtc ggctccctca gcccgggcgg ccgtgggagc ccgcctcgat
     6421 atgtacaccc gagaagctcc cagcgtcctc ctgggccgcg atactcgacc accacgcacg
     6481 cacaccgcac taacgattcg gccggcgctc gattcggccg gcgctcgatt cggccggcgc
     6541 tcgattcggc cggcgctcga ttcggccggc gctcgattcg gccgagcaga agagtgaaca
     6601 accaccgacc acgcttccgc tctgcgcgcc gtacccgacc tacctcccgc agctcgaagc
     6661 agctcccggg agtaccgccg tactcacccg cctgtgctca ccatccaccg acgcaaagcc
     6721 caacccgagc acacctcttg caccaaggtg ccgaccgtgg ctttccgctc gcagggttcc
     6781 agaagaaatc gaacgatcca gcgcggcaag gttcaaaaag caggggttgg tggggaggag
     6841 gttttggggg gtgtcgccgg gatacctgat atggctttgt tttgcgtagt cgaataattt
     6901 tccatatagc ctcggcgcgt cggactcgaa tagttgatgt gggcgggcac agttgcccca
     6961 tgaaatccgc aacggggggc gtgctgagcg atcggcaatg ggcggatgcg gtgttgcttc
     7021 cgcaccggcc gttcgcgacg aacaacctcc aacgaggtca gtaccggatg agccgcgacg
     7081 acgcattggc aatgcggtac gtcgagcatt caccgcacgc gttgctcgga tctatcgtca
     7141 tcgactgcga tcacgttgac gccgcgatgc gcgcattcga gcaaccatcc gaccatccgg
     7201 cgccgaactg ggtcgcacaa tcgccgtccg gccgcgcaca catcggatgg tggctcggcc
     7261 ccaaccacgt gtgccgcacc gacagcgccc gactgacgcc actgcgctac gcccaccgca
     7321 tcgaaaccgg cctcaagatc agcgtcggcg gcgatttcgc gtatggcggg caactgacca
     7381 aaaacccgat tcaccccgat tgggagacga tctacggccc ggccaccccg tacacattgc
     7441 ggcagctggc caccatccac acaccccggc agatgccgcg tcggcccgat cgggccgtgg
     7501 gcctgggccg caacgtcacc atgttcgacg ccacccggcg atgggcatac ccgcagtggt
     7561 ggcaacaccg aaacggaacc ggccgcgact gggaccatct cgtcctgcag cactgccacg
     7621 ccgtcaacac cgagttcacg acaccactgc cgttcaccga agtacgcgcc accgcgcaat
     7681 ccatctccaa atggatctgg cgcaatttca ccgaagaaca gtaccgagcc cgacaagcgc
     7741 atctcggtca aaaaggcggc aaggcaacga cactcgccaa acaagaagcc gtccgaaaca
     7801 atgcaagaaa gtacgacgaa catacgatgc gagaggcgat tatctgatgg gcggagccaa
     7861 aaatccggtg cgccgaaaga tgacggcagc agcagcagcc gaaaaattcg gtgcctccac
     7921 tcgcacaatc caacgcttgt ttgctgagcc gcgtgacgat tacctcggcc gtgcgaaagc
     7981 tcgccgtgac aaagctgtcg agctgcggaa gcaggggttg aagtaccggg aaatcgccga
     8041 agcgatggaa ctctcgaccg ggatcgtcgg ccgattactg cacgacgccc gcaggcacgg
     8101 cgagatttca gcggaggatc tgtcggcgta accaagtcag cgggttgtcg ggttccggcc
     8161 ggcgctcggc actcggaccg gccggcggat ggtgttctgc ctctggcgca gcgtcagcta
     8221 ccgccgaagg cctgtcatcg accggcttcg actgaagtat gagcaacgtc acagcctgtg
     8281 attggatgat ccgctcacgc tcgaccgcta cctgttcagc tgccgcccgc tgggcatgag
     8341 caacggccaa ctctcgttca a
//
