LOCUS       Exported                7641 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pABC5-mRfp1, complete sequence.
ACCESSION   DQ903896
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7641)
  AUTHORS   Lamping E, Monk BC, Niimi K, Holmes AR, Tsao S, Tanabe K, Niimi M, 
            Uehara Y, Cannon RD.
  TITLE     Characterization of three classes of membrane proteins involved in 
            fungal azole resistance by functional hyperexpression in 
            Saccharomyces cerevisiae
  JOURNAL   Eukaryotic Cell 6 (7), 1150-1165 (2007)
  PUBMED    17513564
REFERENCE   2  (bases 1 to 7641)
  AUTHORS   Lamping E, Monk BC, Niimi K.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-AUG-2006) Oral Sciences, Univ. of Otago, 310 Gt. King 
            St, Dunedin, Otago 9001, New Zealand
REFERENCE   3  (bases 1 to 7641)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7641
                     /organism="Cloning vector pABC5-mRfp1"
                     /mol_type="other DNA"
                     /label=derivative of cloning vector pABC5
                     /note="derivative of cloning vector pABC5"
                     /db_xref="taxon:404029"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    687..5399
                     /label=Saccharomyces cerevisiae transformation cassette
                     /note="Saccharomyces cerevisiae transformation cassette"
     regulatory      687..1816
                     /regulatory_class="promoter"
                     /note="Saccharomyces cerevisiae PDR5"
     misc_feature    1817..1854
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    1855..2541
                     /note="Discosoma monomeric red fluorescent protein tag 
                     mRFP1"
     CDS             1855..2532
                     /codon_start=1
                     /gene="mRFP1"
                     /product="monomeric red fluorescent protein"
                     /label=mRFP1
                     /protein_id="ABI82058.1"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     gene            1855..2532
                     /gene="mRFP1"
                     /label=mRFP1
     regulatory      2542..2880
                     /regulatory_class="terminator"
                     /note="Saccharomyces cerevisiae PGK1"
     terminator      2606..2876
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     misc_feature    2887..5131
                     /label=HIS1 selection marker
                     /note="HIS1 selection marker"
     CDS             3860..4753
                     /codon_start=1
                     /gene="HIS1"
                     /product="ATP phosphoribosyltransferase"
                     /label=HIS1
                     /protein_id="ABI82059.1"
                     /translation="MDLVNHLTDRLLFAIPKKGRLYSKSVSILNGADITFHRSQRLDIA
                     LSTSLPVALVFLPAADIPTFVGEGKCDLGITGVDQVRESNVDVDLAIDLQFGNCKLQVQ
                     VPVNGEYKKPEQLIGKTIVTSFVKLAEKYFADLEGTTVEKMTTRIKFVSGSVEASCALG
                     IGDAIVDLVESGETMRAAGLVDIATVLSTSAYLIESKNPKSDKSLIATIKSRIEGVMTA
                     QRFVSCIYNAPEDKLPELLKVTPGRRAPTISKIDDEGWVAVSSMIERKTKGVVLDELKR
                     LGASDIMVFEISNCRV"
     gene            3860..4753
                     /gene="HIS1"
                     /label=HIS1
     misc_feature    5132..5399
                     /label=Saccharomyces cerevisiae PDR5 gene, 3 end
                     /note="Saccharomyces cerevisiae PDR5 gene, 3 end"
     promoter        complement(5453..5471)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5492..5508)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5516..5532
                     /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(5540..5570)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5585..5606
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5894..6482)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6653..7513)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="ABI82060.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7514..7618)
                     /gene="bla"
                     /label=AmpR promoter