LOCUS       40924_44272        9207 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Reporter vector pTRK1-bglT2mcsHEstop DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9207)
  AUTHORS   Vian A, Carrascosa AV, Garcia JL, Cortes E.
  TITLE     Structure of the beta-galactosidase gene from Thermus sp. strain T2:
            expression in Escherichia coli and purification in a single step of 
            an active fusion protein
  JOURNAL   Appl. Environ. Microbiol. 64 (6), 2187-2191 (1998)
  PUBMED    9603833
REFERENCE   2  (bases 1 to 9207)
  AUTHORS   Fujita A, Misumi Y, Koyama Y.
  TITLE     Two versatile shuttle vectors for Thermus thermophilus-Escherichia 
            coli containing multiple cloning sites, lacZalpha gene and kanamycin
            or hygromycin resistance marker
  JOURNAL   Plasmid 67 (3), 272-275 (2012)
  PUBMED    22252135
REFERENCE   3  (bases 1 to 9207)
  AUTHORS   Fujita A, Misumi Y, Honda S, Sato T, Koyama Y.
  TITLE     Construction of new cloning vectors that employ the phytoene 
            synthase encoding gene for color screening of cloned DNA inserts in 
            Thermus thermophilus
  JOURNAL   Gene 527 (2), 655-662 (2013)
  PUBMED    23845779
REFERENCE   4  (bases 1 to 9207)
  AUTHORS   Fujita A, Sato T, Koyama Y, Misumi Y.
  TITLE     Development of a novel reporter gene system incorporating a gene 
            encoding a thermostable beta-galactosidase for use in Thermus 
            thermophilus HB27
  JOURNAL   Unpublished
REFERENCE   5  (bases 1 to 9207)
  AUTHORS   Fujita A.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-JAN-2015) Contact:Atsushi Fujita National Institute of
            Advanced Industrial Science and Technology, Health Research 
            Institute; 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan URL 
            :https://unit.aist.go.jp/hri/
REFERENCE   6  (bases 1 to 9207)
  TITLE     Direct Submission
REFERENCE   7  (bases 1 to 9207)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "1998"; volume: "64"; issue: "6"; pages:
            "2187-2191"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2012"; volume: "67"; issue: "3"; pages: "272-275"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Gene"; 
            date: "2013"; volume: "527"; issue: "2"; pages: "655-662"
COMMENT     SGRef: number: 4; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 5; type: "Journal Article"; journalName: "Submitted 
            (27-JAN-2015) Contact:Atsushi Fujita National Institute of Advanced 
            Industrial Science and Technology, Health Research Institute; 1-8-31
            Midorigaoka, Ikeda, Osaka 563-8577, Japan URL 
            :https://unit.aist.go.jp/hri/"
COMMENT     SGRef: number: 6; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    69..123
                     /note="polylinker 
                     HindIII-NruI-SalI-NotI-PstI-SwaI-EcoRV-PvuII-EcoRI"
     CDS             137..2071
                     /gene="bgaA"
                     /label=bgaA
                     /note="Beta-galactosidase BgaA from Thermus sp. Accession#:
                     O54315"
     CDS             complement(2922..4085)
                     /codon_start=1
                     /gene="repA"
                     /product="putative repA protein"
                     /label=repA
                     /protein_id="BAS05947.1"
                     /translation="MVLRAYAALRGLSPEALRAHLLAPPLRPERAREAFQRPYLAHFAQ
                     TLPRYPYATDDPKEGVRIYKRENALKRVHVQVGHYPHAVLRLVVDVDLPWPQVEERIHA
                     LPPSLVLVNPRSGHFHAWYELDPIPLTPPPGREGSLKGALALLAEVEALLEAYYGADPG
                     YNGLLSRNPFLHPPEWTWGGGKRWSLRDLHRELRGLLPSGTRRRVDPGLASYGRNNALF
                     DRLRAEAYAHVALFRGVPGGEEAFRAWVEQRAHALNQSLFRDHPKGPLDPREVHHTAKS
                     VAKWTYRNYRGARVYPVSSTGRPDRSRLSPQARALIPPLQGQELQEAVREGGRRRGSRR
                     RQEAEEKLTEALKRLQARGERVTARALAREAGVKPHTASKWLKRMRE"
     gene            complement(2922..4085)
                     /gene="repA"
                     /label=repA
     CDS             6577..7347
                     /codon_start=1
                     /gene="KNT"
                     /product="kanamycin nucleotidyltransferase"
                     /label=KNT
                     /protein_id="BAS05948.1"
                     /translation="MRIVNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGR
                     QTDGPYSDIEMMCVMSTEEAEFSHEWTTGEWKAEVNFYSEEILLDYASRVEPDWPLTHG
                     RFFSILPIYDPGGYFEKVYQTAKSVEAQKFHDAICALIVEELFEYAGKWRNIRVQGPTT
                     FLPSLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQPDLPPGYVQLCQLVMSGQLSD
                     PEKLLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     gene            6577..7347
                     /gene="KNT"
                     /label=KNT
     CDS             7406..8263
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8437..9025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagccatat
       61 gaagaattaa gcttcgcgag tcgacgcggc cgcctgcaga tttaaatgat atcagctgaa
      121 ttcttgaatg agtcgtatgt tgggcgtttg ttactacccg gagcattggc ctgaagaacg
      181 ttgggaagag gactttaagg ccatgcgggc tctggggctc cgctatgtgc gcctggggga
      241 gtttgcctgg agcgccctcg agcccacccc aggtgctctc cgctggggtt ggctggatcg
      301 ggttctggat ctggcgcaga aggagggcct agccgtagtc ctcggtaccc ccaccgctac
      361 cccccccaag tggttggtgg accggtaccc ggagatcctc cctgtggacc gggaggggcg
      421 gaggcggaac tttggtgggc ggcggcacta ctgcttctcc agccccgcct accgggaaga
      481 gactgcccgc atcgtggccc ttctagcgga gcgttacggc cgccaccccg cggtggtagg
      541 gttccaggtg gacaacgagt ttggctgtca cggcaccgtg cgctgctact gtcccaactg
      601 tcgtgaggcc ttccggggct ggttgagggc taagtacggg acgattgacg cgctgaacgc
      661 cgcctgggga acggtctttt ggagccaaac gtaccgggat ttcggtgagg tggagcttcc
      721 tcacctcacc gtggccgagg ccaaccccag ccaccttctg gactactacc gtttcgcatc
      781 agaccaggtg cgggcctaca atcgtttcca agtggacctc cttcgggata atgctcctgg
      841 ccggttcatt acccataact tcatggggtt tttcaccgat ctggacccct ttgccttggc
      901 cgaggacctg gattttgccg cctgggacag ctaccccttg ggcttcaccg acctgatgcc
      961 ccttccccag gaggagaagg ttcagtgggc ccgcacgggc caccccgatg tggccgcctt
     1021 ccaccacgac ctttaccggg gggtagggcg ggggcggttt tgggtcatgg agcagcagcc
     1081 gggtcccgtg aactgggccc cccacaatcc aagcccggcg cccgggatgg tacgcctttg
     1141 gacttgggag gccatagccc atggggcaga ggtggtttcc tacttccgct ggcgccaagc
     1201 gccctttgcc caagagcaga tgcaagcggg gttcaaccgt ccagatttcc agccggaagt
     1261 ggcctttttt gaagtgcagc gcgtagcaga ggagctttcc gccctccccc ttcctcccgc
     1321 agggtgtgcc cccgtggctt tggtttatga ttccgaggcg gcttgggtgt ttgagatcca
     1381 gccccaaggg gccgagtgga aatacctgac cctggtgttt tccttttata gcgttttccg
     1441 gcgcctaggg ttggaagtgg acatttttaa gcccggggca gaattgggcg ggtatggcct
     1501 ggtggtggtt cccagcctgc ccattgtgcg taaggaggcc ctcgaggccc tttcccaggc
     1561 ggacggcctg gtaattgtcg gaccccgttc ggggagcaaa acggagaaat tccagatccc
     1621 ccccgaaatc cctccgggcg cgctccaagc cctccttccc cttaaggtag tgcgggtgga
     1681 gagcctgccc ccggggcttt tggaggaggc cgaaggaccc tggggccgct ttgccttcgg
     1741 ggtttggcgg gagtgggtgg agaccgatct tccccccttg ctccgtttta cggatggtgg
     1801 gggaatcctt ttccgcaggg gtcgctacct gtacctggcg gcttggccca gcccagaact
     1861 tctctttgcc ctttgccagt ccttggccga ggaggcgggc ttgcatcccc gcttcctccc
     1921 tgagggcctg cgtttacgga gacggggccc gttggtgttt gcctttaact atggccccga
     1981 ggtggtggag gcacctgccc ctccaggggt gcggtttctc ttgggggata ggcgtattcc
     2041 cccccatgac cttgccgtgt gggaggagac atgaggcttg tactgggcgg cttggctggc
     2101 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     2161 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     2221 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     2281 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     2341 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     2401 cgagacgaaa gggcggctat ggaggggttc tccctgtcta cgcttgagcg cccgagggcc
     2461 ggtccagccc ccgggccaga gcgggctccg gaccctgccc ccagaaggac cctctcaccc
     2521 gtcctaaggc ccctagaagg ccggggggcg gggggagggg tgggattggt gccccccgcc
     2581 caccccccgc cttaaaagcc gttttagggg cggtcctcga gggggaaaat cagtgttttt
     2641 gccgtcctgg acgatgaaaa ttgcgtgcgg gcggggtaaa tatgtgagtt atctcacttt
     2701 ctctcggatc tcaaggtgtc taccccaaga aaaacaaaat ttttgcggct tccaggaaaa
     2761 agaaggggga ctttagccgt caactacgct aaggagactc tctaggttct ccaatacagc
     2821 cccttcgggg ctggcggggg ttgcaccccc gcacccccgc ctgtataccc tgatagccac
     2881 cagcttgggg cacatttttg gggtttgtgc cgtcctggac gctactcccg catcctcttc
     2941 aaccacttgg aggcggtatg gggcttgacc cccgcctccc gggccagggc cctggccgtg
     3001 acccgctccc cccgggcctg gaggcgcttc agggcctccg tgagcttctc ctcggcctcc
     3061 tgcctgcgcc gggatccgcg ccgccttccg ccctcccgca ccgcctcctg gagctcctgg
     3121 ccctggaggg gcgggatcag ggcccgggcc tggggagaga ggcggctccg gtccggcctc
     3181 cccgtggagg agaccgggta gaccctcgcc ccccggtagt tccggtaggt ccacttggcc
     3241 acgctcttcg ccgtgtggtg gacctcccgg gggtcaaggg gccccttggg gtggtcccgg
     3301 aagagggact ggttcagggc gtgggccctc tgctccaccc aggcccggaa ggcctcctcc
     3361 cccccgggga cgccccggaa gagggccacg tgggcgtagg cctccgcccg caggcggtca
     3421 aacagggcgt tgttccgccc gtaggaggcc aggcccgggt ccacccgcct ccgggtcccg
     3481 gagggaagga gcccccggag ctcccggtgg aggtcccgca ggctccaccg cttccccccg
     3541 ccccaggtcc actccggggg gtggaggaag gggtttcggg agaggagacc gttgtagccc
     3601 gggtccgccc cgtagtaggc ctccagcagg gcctccacct ccgcgagaag ggccagggcc
     3661 cccttcaggc tcccctcccg cccgggcggg ggcgtgaggg ggatggggtc cagctcgtac
     3721 caggcgtgga agtggcccga tctcgggttg accaggacca gggagggggg gagggcgtgg
     3781 atccgctcct ccacctgggg ccaggggagg tccacgtcca ccaccagccg caagacggcg
     3841 tgggggtagt ggcccacctg gacgtggacc cgcttcaggg cgttctcccg cttgtagatg
     3901 cgcaccccct ccttggggtc gtccgtggcg taggggtagc gggggagagt ctgggcgaag
     3961 tgggcgaggt agggccgctg gaaggcctcc cgggcccgct ccgggcggag gggaggggcc
     4021 aggaggtggg cgcggagggc ctccggggag aggccgcgca gggcggcgta ggcgcgaaga
     4081 accacctccc ccaggtcggg gtttatctgg aagagtcctg cgattttagc gagggtgtcc
     4141 gggatttgcg cttccggccg ggtgtggtgc tccatctctt gcctccttcc ccccggccgg
     4201 gttaggatgg aggcctcctg gggaggagag gcctcggcgg ttaacccggc cgaggtctgt
     4261 ttcattcatg ccccccattc tagcgacaag ccccgggacc aagcggtagt ccccaggtca
     4321 atgcccccag aaccgccacc agggccgcct ccgcgggggc tccagggcct tcagggcctc
     4381 ccggaggcgg gcgttctcct cccgcagggc ccttagctcc ccctccaccc gctccagccg
     4441 ctcggccagg gcccggagga gggccagggc ctccccctcg gaggggaggg cgagggccgg
     4501 gaagtctccc tggacccggg ccagggcctc ctccaggggg aggccttccc gcaggtgggc
     4561 ctcccgggcc gcccggaggc gggccagggc ctccttgggc cagagccgtc ccccccgggg
     4621 atcccggggc aggggaccta ccaaccgctc ccacagggcg gcgtagcgcc ggagggtggc
     4681 cggggagacc cccagggccc gggcggccaa ggccgggggc aggagggtcg ggtcttcccc
     4741 ttccgggtgc tccacgaccc cagtctaccc aggcgctcag ggtgagcggc ctcgtgatac
     4801 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4861 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4921 atccgctcat gagacaataa ccctgataaa tgcttcaata atatctttgt tcttactccc
     4981 atagatcttc atgtgttgta ctacccccag aatcccagcc tctgccccgg aggagcggga
     5041 gcggtgagca ccaacaaccc agggttctac agcctgcccc tgaagcccac cacctcccct
     5101 cccacgcccc cgggaccggg ggagatcatt ccttttccga ctcctagtta tgggcccgtc
     5161 acgctgctac ctttctacac gagcgaccgc ccagagaagt acgacctgac cttctactgc
     5221 aacgccgggg aggggccgcc acctgaccac ccccggcctg gtgggccagc atcctggatc
     5281 ctggcgagga ggtgggcgtt gcgggccaac gggcctatcc gtggcggcga ccgccagccg
     5341 gcgtcccaaa acgccagggg gcgacagggc cgagaagggg ttgaccggga ggtcggcctg
     5401 gccgagaagg tcgccccgag agggcgcagc gggagtcgcc gtggtccccc ggcctcacag
     5461 ggggggaagg gaactgccag agtgccggct agccgcgggc ctcaacgcgg tggtggccac
     5521 gggcctaacc gtggcggcga aggccggcca aacgggccag ccacggcgtc ctccggccgt
     5581 acaggagggg gcgttcccgg gcccagaaac cgggagatgg gaagcccgga gctccccgcg
     5641 gggggttccc gggcggtggc cggaagggcc tgcgggcccg gaaggacacg aatctcaggg
     5701 cgagaagccc acccttacag ggggacaccc ttatccccca cccccggctg acacctcagc
     5761 cggggttttc tgtttgcccc tcaagccgcc tggaaggccc ctggaaggcc ggggaagggg
     5821 tggggggcta cttccccttg gccctcgagg gggcggcctt ctagggccct tagaggccgg
     5881 gagagggggc cccggggagc tccgagggag gaagaggcag gcgcaggacc gcctccccca
     5941 aaagcctccc ccgctccacc acccccagga ccaggaccaa gccccaggag gcggggagga
     6001 ggtggggcct ccgggccatg aagcgcacca caaggggcct caccgggaag ccgcacccgc
     6061 gcccccttcc atggctcacc aaattctcat gttgggagag caacagttag ggtatgggtg
     6121 gaatcaatca caaaccgacc aaaaagataa tgctcgtttc ttccgcagct cgtgtgagtc
     6181 gggtcgttgg tatgacgctt gaaacccttc tcgctgcgaa caccgcatat gaggaagcca
     6241 tcattgcagc atttaacggt tcgtacgacg aggcactttc tcaccacaac cagagcctgg
     6301 accttgcggc aaccgcaaag cagcgcttgc gcgacattga tgctgcttac accatgatgc
     6361 tcgaagacat cgctattgag aaatatccgg gtaaccgctg gctccaaagc tcgaacctga
     6421 tgtgcttcgc aaagagcttc aggaaaggtg ctacttgatt gacacagtct ctttgatgaa
     6481 atggccagtg cgatccacgg ctggtggaaa gaaaaacagg cagaacagcg gtgatcccgt
     6541 cgagctcgag caaatctaaa attatctgaa aagggaatga gaatagtgaa tggaccaata
     6601 ataatgacta gagaagaaag aatgaagatt gttcatgaaa ttaaggaacg aatattggat
     6661 aaatatgggg atgatgttaa ggctattggt gtttatggct ctcttggtcg tcagactgat
     6721 gggccctatt cggatattga gatgatgtgt gtcatgtcaa cagaggaagc agagttcagc
     6781 catgaatgga caaccggtga gtggaaggcg gaagtgaatt tttatagcga agagattcta
     6841 ctagattatg catctcgggt ggaaccggat tggccgctta cacatggtcg atttttctct
     6901 attttgccga tttatgatcc aggtggatac tttgagaaag tgtaccaaac tgctaaatcg
     6961 gtagaagccc aaaagttcca cgatgcgatc tgtgccctta tcgtagaaga gctgtttgaa
     7021 tatgcaggca aatggcgtaa tattcgtgtg caaggaccga caacatttct accatccttg
     7081 actgtacagg tggcaatggc aggtgccatg ttgattggtc tgcatcatcg catctgttat
     7141 acgacgagcg cttcggtctt aactgaagca gttaagcaac cagatcttcc tccaggttat
     7201 gtccaactgt gccagctcgt aatgtctggt caactttccg accctgagaa acttctggaa
     7261 tcgctagaga atttctggaa tgggattcag gagtggacag aacgacacgg atatatagtg
     7321 gatgtgtcaa aacgcatacc attttgaacg atgacctcta ataattgtta atcatgttgg
     7381 ttacggatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     7441 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     7501 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     7561 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     7621 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     7681 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     7741 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     7801 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     7861 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     7921 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     7981 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     8041 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     8101 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     8161 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     8221 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     8281 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     8341 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     8401 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     8461 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     8521 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     8581 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     8641 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     8701 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     8761 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     8821 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     8881 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     8941 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     9001 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     9061 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     9121 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     9181 cgagtcagtg agcgaggaag cggaaga
//
