LOCUS       Exported                4871 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC18T-mini-Tn7T-aadA, complete sequence.
ACCESSION   KU936038
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4871)
  AUTHORS   Lehman SS, Mladinich K, Boonyakanog A, Mima T, Karkhoff-Schweizer 
            RR, Schweizer HP.
  TITLE     Versatile nourseothricin and streptomycin/spectinomycin resistance 
            gene cassettes and their use in chromosome integration vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4871)
  AUTHORS   Lehman SS, Mladinich K, Boonyakanog A, Mima T, Karkhoff-Schweizer 
            RR, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-MAR-2016) Molecular Genetics and Microbiology, 
            University of Florida, 2055 Mowry Road, Gainesville, FL 32610, USA
REFERENCE   3  (bases 1 to 4871)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4871
                     /organism="Cloning vector pUC18T-mini-Tn7T-aadA"
                     /mol_type="other DNA"
                     /db_xref="taxon:1884653"
     mobile_element  382..580
                     /mobile_element_type="transposon:Tn7R"
                     /note="right end of Tn7"
     primer_bind     585..601
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      629..723
                     /regulatory_class="terminator"
                     /note="bacteriophage lambda T0"
     terminator      629..723
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      820..917
                     /regulatory_class="terminator"
                     /note="Escherichia coli rrnB T1"
     terminator      826..912
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    955..988
                     /note="loxP; Cre recombinase site"
     CDS             complement(1159..1950)
                     /codon_start=1
                     /gene="aadA"
                     /product="streptomycin/spectinomycin adenyltransferase"
                     /label=aadA
                     /protein_id="ANY60778.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            complement(1159..1950)
                     /gene="aadA"
                     /label=aadA
     regulatory      2052..2101
                     /regulatory_class="promoter"
                     /note="aadA gene promoter"
     misc_feature    2254..2287
                     /note="loxP; Cre recombinase site"
     protein_bind    2254..2287
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2302..2376
                     /note="MCS; multiple cloning site"
     mobile_element  2388..2553
                     /mobile_element_type="transposon:Tn7L"
                     /note="left end of Tn7"
     oriT            2705..2927
     oriT            2712..2820
                     /note="incP origin of transfer"
     rep_origin      2975..3643
                     /label=pMB
                     /note="pMB"
     rep_origin      complement(3052..3640)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3811..4671)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="ANY60779.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3811..4671)
                     /gene="bla"
                     /label=bla
     promoter        complement(4672..4776)
                     /gene="bla"
                     /label=AmpR promoter