LOCUS       Exported                9568 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Insertion vector pUTTnsKm, complete sequence.
ACCESSION   GQ371204
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9568)
  AUTHORS   Li R, Wang G, Shen B, Wang R, Song Y, Li S, Jiang J.
  TITLE     Random transposon vectors pUTTns for the markerless integration of 
            exogenous genes into gram-negative eubacteria chromosomes
  JOURNAL   J. Microbiol. Methods 79 (2), 220-226 (2009)
  PUBMED    19778558
REFERENCE   2  (bases 1 to 9568)
  AUTHORS   Li R, Jiang J, Li S.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-JUL-2009) College of Life Science, Nanjing 
            Agricultural University, 6# Tongwei Road, Nanjing, Jiangsu 210095, 
            China
REFERENCE   3  (bases 1 to 9568)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9568
                     /organism="Insertion vector pUTTnsKm"
                     /mol_type="other DNA"
                     /db_xref="taxon:665876"
     CDS             complement(311..1171)
                     /codon_start=1
                     /gene="bla"
                     /product="AmpR"
                     /label=bla
                     /protein_id="ACU42196.1"
                     /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
                     /protein_id="ACU42197.1"
                     /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(3253..4674)
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase"
                     /label=sacB
                     /protein_id="ACU42198.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            complement(3253..4674)
                     /gene="sacB"
                     /label=sacB
     CDS             5465..6280
                     /codon_start=1
                     /gene="km"
                     /product="KanR"
                     /label=km
                     /note="kanamycin resistance protein"
                     /protein_id="ACU42199.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            5465..6280
                     /gene="km"
                     /label=km
     misc_feature    6739..7170
                     /label=similar to sacB
                     /note="similar to sacB"
     misc_feature    7171..7200
                     /label=MCS
                     /note="MCS"
     misc_feature    7201..7219
                     /label=Tn5 OE end
                     /note="Tn5 OE end"
     protein_bind    7231..7247
                     /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)."
     misc_feature    7279..7666
                     /note="oriRR6K alpha peptide; replicate origin"
     rep_origin      7282..7670
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    7696..9424
                     /note="mob RP4; similar to Escherichia coli sequence 
                     deposited in INSD Accession X54459"
     CDS             complement(8288..8659)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            8692..8801
                     /note="incP origin of transfer"