LOCUS       Exported                8296 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Transposon delivery vector pUT-miniTn5-gfp-tet, complete sequence.
ACCESSION   AY364166
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8296)
  AUTHORS   Schuler L, Wattiau P.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-AUG-2003) Bioengineering Unit, UCL, Place Croix du Sud
            2 Pbox 19, Louvain-la-Neuve 1348, Belgium
REFERENCE   2  (bases 1 to 8296)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8296
                     /organism="Transposon delivery vector pUT-miniTn5-gfp-tet"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:243674"
     CDS             complement(311..1171)
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /label=bla
                     /note="beta-lactamase from Tn3"
                     /protein_id="AAQ67239.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(311..1171)
                     /gene="bla"
                     /label=bla
     promoter        complement(1172..1276)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(1323..2753)
                     /codon_start=1
                     /gene="tnpA"
                     /product="TnpA"
                     /label=tnpA
                     /note="Tn5 transposase"
                     /protein_id="AAQ67236.1"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSEAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTLPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     gene            complement(1323..2753)
                     /gene="tnpA"
                     /label=tnpA
     misc_feature    2871..2889
                     /label=Tn5 IE end
                     /note="Tn5 IE end"
     CDS             complement(3356..4546)
                     /codon_start=1
                     /gene="tetA"
                     /product="TetA"
                     /label=tetA
                     /note="class C tetracyclin resistance protein"
                     /protein_id="AAQ67237.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            complement(3356..4546)
                     /gene="tetA"
                     /label=tetA
     primer_bind     5107..5123
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(5154..5870)
                     /codon_start=1
                     /gene="gfp"
                     /allele="mut2"
                     /product="Gfp-mut2"
                     /label=gfp
                     /note="mutant green fluorescent protein"
                     /protein_id="AAQ67238.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            complement(5154..5870)
                     /gene="gfp"
                     /allele="mut2"
                     /label=gfp
     misc_feature    5929..5947
                     /label=Tn5 OE end
                     /note="Tn5 OE end"
     protein_bind    5959..5975
                     /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      6010..6398
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    6890..7663
                     /label=RK2/RP4 mobilization origin
                     /note="RK2/RP4 mobilization origin"
     CDS             complement(7016..7387)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            7420..7529
                     /note="incP origin of transfer"