LOCUS       Exported                5935 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Mini Tn5 transposon vector pUT-dfrA17, complete sequence.
ACCESSION   KF765532
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5935)
  AUTHORS   Tsai Y, Chu C, Lee C-H., Liaw S-J.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-OCT-2013) Department of Clinical Laboratory Sciences 
            and Medical Biotechnology, College of Medicine, National Taiwan 
            University, No. 1, Chang-Te St., Taipei 100, Taiwan
REFERENCE   2  (bases 1 to 5935)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5935
                     /organism="mini Tn5 transposon vector pUT-dfrA17"
                     /host="Escherichia coli S17-1"
                     /mol_type="other DNA"
                     /country="Taiwan"
                     /PCR_primers="fwd_name: dfra17-f, fwd_seq:
                     ggtacctcatggcttgttatgactgt, rev_name: dfra17-r, rev_seq: 
                     gcggccgcattgttagccttttt"
                     /PCR_primers="fwd_name: blatem-f, fwd_seq:
                     gattgtactgagagtgcac, rev_name: blatem-r, rev_seq: 
                     cccctatttgtttatttttc"
                     /note="PCR_primers=rev_name: i-out primer, rev_seq: 
                     gagctcgaattcggcctag"
                     /db_xref="taxon:1448878"
     CDS             complement(311..1171)
                     /codon_start=1
                     /gene="blaTEM"
                     /product="beta-lactamase"
                     /label=blaTEM
                     /note="BlaTEM"
                     /protein_id="AHG52770.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(311..1171)
                     /gene="blaTEM"
                     /label=blaTEM
     promoter        complement(1172..1276)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(1323..2753)
                     /codon_start=1
                     /gene="tnpA"
                     /product="transposase TnpA"
                     /label=tnpA
                     /protein_id="AHG52771.1"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSEAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTLPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     gene            complement(1323..2753)
                     /gene="tnpA"
                     /label=tnpA
     misc_feature    2871..2889
                     /note="Tn5 inside end; IE"
     CDS             3007..3555
                     /codon_start=1
                     /gene="dfrA17"
                     /product="dihydrofolate reductase DfrA17"
                     /label=dfrA17
                     /protein_id="AHG52772.1"
                     /translation="MLWSSNDVTQQGSRPKTKLAIKGVKLKISLISAVSENGVIGSGPD
                     IPWSVKGEQLLFKALTYNQWLLVGRKTFDSMGVLPNRKYAVVSKNGISSSNENVLVFPS
                     IENALKELSKVTDHVYVSGGGQIYNSLIEKADIIHLSTVHVEVEGDIKFPIMPENFNLV
                     FEQFFMSNINYTYQIWKKG"
     gene            3007..3555
                     /gene="dfrA17"
                     /label=dfrA17
     misc_feature    3568..3586
                     /note="Tn5 outside end; OE"
     protein_bind    3598..3614
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     rep_origin      3646..4033
                     /label=oriRR6K alpha peptide
                     /note="oriRR6K alpha peptide"
     rep_origin      3649..4037
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    4529..5302
                     /label=RK2/RP4 mobilization origin
                     /note="RK2/RP4 mobilization origin"
     CDS             complement(4655..5026)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            5059..5168
                     /note="incP origin of transfer"