LOCUS       Exported                9207 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Reporter vector pTRK1-bglT2mcsHEstop DNA, complete sequence.
ACCESSION   LC021516
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9207
                     /organism="Reporter vector pTRK1-bglT2mcsHEstop"
                     /mol_type="other DNA"
                     /note="a reporter vector for Thremus thermophilus HB27, 
                     pTRK1-bglT2mcsHEstop"
                     /db_xref="taxon:1607993"
     misc_feature    69..123
                     /note="polylinker 
                     HindIII-NruI-SalI-NotI-PstI-SwaI-EcoRV-PvuII-EcoRI"
     CDS             137..2074
                     /codon_start=1
                     /product="beta-galactosidase"
                     /label=beta-galactosidase
                     /note="Thermus spp T2 beta-galactosidase"
                     /protein_id="BAS05946.1"
                     /translation="MLGVCYYPEHWPEERWEEDFKAMRALGLRYVRLGEFAWSALEPTP
                     GALRWGWLDRVLDLAQKEGLAVVLGTPTATPPKWLVDRYPEILPVDREGRRRNFGGRRH
                     YCFSSPAYREETARIVALLAERYGRHPAVVGFQVDNEFGCHGTVRCYCPNCREAFRGWL
                     RAKYGTIDALNAAWGTVFWSQTYRDFGEVELPHLTVAEANPSHLLDYYRFASDQVRAYN
                     RFQVDLLRDNAPGRFITHNFMGFFTDLDPFALAEDLDFAAWDSYPLGFTDLMPLPQEEK
                     VQWARTGHPDVAAFHHDLYRGVGRGRFWVMEQQPGPVNWAPHNPSPAPGMVRLWTWEAI
                     AHGAEVVSYFRWRQAPFAQEQMQAGFNRPDFQPEVAFFEVQRVAEELSALPLPPAGCAP
                     VALVYDSEAAWVFEIQPQGAEWKYLTLVFSFYSVFRRLGLEVDIFKPGAELGGYGLVVV
                     PSLPIVRKEALEALSQADGLVIVGPRSGSKTEKFQIPPEIPPGALQALLPLKVVRVESL
                     PPGLLEEAEGPWGRFAFGVWREWVETDLPPLLRFTDGGGILFRRGRYLYLAAWPSPELL
                     FALCQSLAEEAGLHPRFLPEGLRLRRRGPLVFAFNYGPEVVEAPAPPGVRFLLGDRRIP
                     PHDLAVWEET"
     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..8266
                     /codon_start=1
                     /gene="Amp-R"
                     /product="beta-lactamase"
                     /label=Amp-R
                     /protein_id="BAS05949.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            7406..8266
                     /gene="Amp-R"
                     /label=Amp-R
     rep_origin      8437..9025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"