LOCUS       Exported                4838 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC18T-mini-Tn7T-nat, complete sequence.
ACCESSION   KU936037
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4838)
  AUTHORS   Lehman SS, Mladinich KM, 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   J. Microbiol. Methods 129, 8-13 (2016)
  PUBMED    27457407
REFERENCE   2  (bases 1 to 4838)
  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 4838)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4838
                     /organism="Cloning vector pUC18T-mini-Tn7T-nat"
                     /mol_type="other DNA"
                     /db_xref="taxon:1884654"
     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    965..987
                     /note="loxP; Cre recombinase site"
     promoter        1117..1460
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     regulatory      1235..1284
                     /regulatory_class="promoter"
                     /note="nat gene promoter"
     CDS             1461..2033
                     /codon_start=1
                     /gene="nat"
                     /product="nourseothricin N-acetyltransferase"
                     /label=nat
                     /protein_id="ANY60776.1"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     gene            1461..2033
                     /gene="nat"
                     /label=nat
     misc_feature    2210..2243
                     /note="loxP; Cre recombinase site"
     protein_bind    2210..2243
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2269..2343
                     /note="MCS; multiple cloning site"
     mobile_element  2355..2520
                     /mobile_element_type="transposon:Tn7L"
                     /note="left end of Tn7"
     oriT            2672..2939
     oriT            2679..2787
                     /note="incP origin of transfer"
     rep_origin      2942..3610
                     /label=pMB
                     /note="pMB"
     rep_origin      complement(3019..3607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3778..4638)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="ANY60777.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3778..4638)
                     /gene="bla"
                     /label=bla
     promoter        complement(4639..4743)
                     /gene="bla"
                     /label=AmpR promoter