LOCUS       Exported                8750 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pTRK1-PRslpA-42MCS DNA, complete sequence.
ACCESSION   LC310789
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8750)
  AUTHORS   Fujita A, Misumi Y.
  TITLE     Development of a new host-vector system for colour selection of 
            cloned DNA inserts using a newly designed beta-galactosidase gene 
            containing multiple cloning sites in Thermus thermophilus HB27
  JOURNAL   Extremophiles 21 (6), 1111-1117 (2017)
  PUBMED    28894938
REFERENCE   2  (bases 1 to 8750)
  AUTHORS   Fujita A.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-JUL-2017) Contact:Atsushi Fujita Institute of Advanced
            Science and Technology, Biomedical Research Institute; 1-8-31 
            Midorigaoka, Ikeda, Osaka 563-8577, Japan URL 
            :https://unit.aist.go.jp/bmd/
REFERENCE   3  (bases 1 to 8750)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8750
                     /organism="Shuttle vector pTRK1-PRslpA-42MCS"
                     /mol_type="other DNA"
                     /db_xref="taxon:2020290"
     regulatory      168..667
                     /regulatory_class="promoter"
                     /note="slpA promoter"
     CDS             683..1978
                     /codon_start=1
                     /gene="the designed TTP0042 gene"
                     /product="beta-galactosidase"
                     /label=the designed TTP0042 gene
                     /protein_id="BBB87064.1"
                     /translation="MTENAEKFLWGVATSAYQIEGATQEDGRGPSIWDAFAQRPGAIRD
                     GSTGEPACDHYRRYEEDIALMQSLGVRAYRFSVAWPRILPEGRGRINPKGLAFYDRLVD
                     RLLASGITPFLTLYHWDLPLALEERGGWRSRETAFAFAEYAEAVARALADRVPFFATLN
                     EPWCSAFLGHWTGEHAPGLRNLEAALRAAHHLLLGHGLAVEALRAAGARRVGIVLNFAP
                     AYGEDPEAVDVADRYHNRFFLDPILGKGYPESPFRDPPPVPILSRDLELVARPLDFLGV
                     NYYAPVRVAPGTGTLPVRYLPPEGPATAMGWEVYPEGLHHLLKRLGREVPWPLYVTENG
                     AAYPDLWTGEAVVEDPERVAYLEAHVEAALRAREEGVDLRGYFVWSLMDNFEWAFGYTR
                     RFGLYYVDFPSQRRIPKRSALWYRERIARAQT"
     gene            683..1978
                     /gene="the designed TTP0042 gene"
                     /label=the designed TTP0042 gene
     misc_feature    791..797
                     /gene="the designed TTP0042 gene"
                     /label=cloning site BlpI
                     /note="cloning site BlpI"
     misc_feature    836..841
                     /gene="the designed TTP0042 gene"
                     /label=cloning site SphI
                     /note="cloning site SphI"
     misc_feature    866..871
                     /gene="the designed TTP0042 gene"
                     /label=cloning site EcoRV
                     /note="cloning site EcoRV"
     misc_feature    883..888
                     /gene="the designed TTP0042 gene"
                     /label=cloning site HindIII
                     /note="cloning site HindIII"
     misc_feature    895..900
                     /gene="the designed TTP0042 gene"
                     /label=cloning site BssHII
                     /note="cloning site BssHII"
     misc_feature    927..932
                     /gene="the designed TTP0042 gene"
                     /label=cloning site EcoRI
                     /note="cloning site EcoRI"
     misc_feature    985..990
                     /gene="the designed TTP0042 gene"
                     /label=cloning site SpeI
                     /note="cloning site SpeI"
     misc_feature    989..994
                     /gene="the designed TTP0042 gene"
                     /label=cloning site SalI
                     /note="cloning site SalI"
     misc_feature    1002..1007
                     /gene="the designed TTP0042 gene"
                     /label=cloning site NruI
                     /note="cloning site NruI"
     misc_feature    1082..1087
                     /gene="the designed TTP0042 gene"
                     /label=cloning site XbaI
                     /note="cloning site XbaI"
     misc_feature    1192..1197
                     /gene="the designed TTP0042 gene"
                     /label=cloning site AvrII
                     /note="cloning site AvrII"
     misc_feature    1944..1949
                     /gene="the designed TTP0042 gene"
                     /label=cloning site Acc65I
                     /note="cloning site Acc65I"
     CDS             6120..6890
                     /codon_start=1
                     /gene="KNT"
                     /product="kanamycin nucleotidiltransferase"
                     /label=KNT
                     /protein_id="BBB87065.1"
                     /translation="MRIVNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGR
                     QTDGPYSDIEMMCVMSTEEAEFSHEWTTGEWKAEVNFYSEEILLDYASRVEPDWPLTHG
                     RFFSILPIYDPGGYFEKVYQTAKSVEAQKFHDAICALIVEELFEYAGKWRNIRVQGPTT
                     FLPSLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQPDLPPGYVQLCQLVMSGQLSD
                     PEKLLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     gene            6120..6890
                     /gene="KNT"
                     /label=KNT
     CDS             6949..7809
                     /codon_start=1
                     /gene="Amp-R"
                     /product="beta-lactamase"
                     /label=Amp-R
                     /protein_id="BBB87066.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            6949..7809
                     /gene="Amp-R"
                     /label=Amp-R
     rep_origin      7980..8568
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"