LOCUS       Exported                4324 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Reporter vector pUS23, complete sequence.
ACCESSION   AY894753
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4324)
  AUTHORS   Coleman NV, Holmes AJ.
  TITLE     The native Pseudomonas stutzeri strain Q chromosomal integron can 
            capture and express cassette-associated genes
  JOURNAL   Microbiology (Reading, Engl.) 151 (PT 6), 1853-1864 (2005)
  PUBMED    15941993
REFERENCE   2  (bases 1 to 4324)
  AUTHORS   Coleman NV, Holmes AJ.
  TITLE     Capture and expression of an aadB-gfp reporter plasmid by the 
            Pseudomonas stutzeri strain Q chromosomal integron
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 4324)
  AUTHORS   Coleman NV, Holmes AJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JAN-2005) School of Molecular and Microbial 
            Biosciences, University of Sydney, Building G08, Sydney, NSW 2006, 
            Australia
REFERENCE   4  (bases 1 to 4324)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4324
                     /organism="Reporter vector pUS23"
                     /mol_type="other DNA"
                     /label=derived from Cloning vector pUC19
                     /note="derived from Cloning vector pUC19"
                     /db_xref="taxon:313917"
     misc_feature    1..33
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      34..288
                     /regulatory_class="terminator"
                     /note="rrnB T1 and T2"
     terminator      39..124
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      216..243
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_recomb     325..384
                     /label=aadB 59-base element (attC) recombination site
                     /note="aadB 59-base element (attC) recombination site"
     RBS             402..410
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     gene            403..948
                     /gene="aadB"
                     /label=aadB
     regulatory      403..409
                     /regulatory_class="ribosome_binding_site"
                     /gene="aadB"
     CDS             415..948
                     /codon_start=1
                     /gene="aadB"
                     /product="aminoglycoside 2-adenylyltransferase"
                     /label=aadB
                     /note="AadB; confers resistance to gentamicin, kanamycin, 
                     and tobramycin"
                     /protein_id="AAX14064.1"
                     /translation="MDTTQVTLIHKILAAADERNLPLWIGGGWAIDARLGRVTRKHDDI
                     DLTFPGERRGELEAIVEMLGGRVMEELDYGFLAEIGDELLDCEPAWWADEAYEIAEAPQ
                     GSCPEAAEGVIAGRPVRCNSWEAIIWDYFYYADEVPPVDWPTKHIESYRLACTSLGAEK
                     VEVLRAAFRSRYAA"
     gene            955..1684
                     /gene="gfpmut3"
                     /label=gfpmut3
     regulatory      955..961
                     /regulatory_class="ribosome_binding_site"
                     /gene="gfpmut3"
     CDS             968..1684
                     /codon_start=1
                     /gene="gfpmut3"
                     /product="green fluorescent protein mut3 variant"
                     /label=gfpmut3
                     /note="Gfpmut3"
                     /protein_id="AAX14065.1"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1708..1724)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1732..1748
                     /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)."
     promoter        complement(1756..1786)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     regulatory      complement(1757..1786)
                     /regulatory_class="promoter"
                     /note="lac"
     protein_bind    1801..1822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2109..2717
                     /label=from pUC19
                     /note="from pUC19"
     misc_RNA        complement(2109..2662)
                     /product="RNAII"
                     /label=replication primer
                     /note="replication primer"
     rep_origin      complement(2110..2698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      2516..2545
                     /regulatory_class="promoter"
                     /note="RNAI"
     misc_RNA        2554..2649
                     /product="RNAI"
                     /label=antisense repressor
                     /note="antisense repressor"
     regulatory      complement(2722..2759)
                     /regulatory_class="promoter"
                     /note="RNAII"
     repeat_region   2722..2759
                     /label=Tn3 terminal inverted repeat
                     /note="Tn3 terminal inverted repeat"
     gene            complement(2869..3799)
                     /gene="bla"
                     /label=bla
     CDS             complement(2869..3729)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="Bla; confers resistance to ampicillin"
                     /protein_id="AAX14066.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3730..3834)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(3733..3739)
                     /regulatory_class="ribosome_binding_site"
                     /gene="bla"
     regulatory      complement(3771..3799)
                     /regulatory_class="promoter"
                     /gene="bla"
     primer_bind     4308..4324
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"