LOCUS       Exported                5824 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pABC3-FlagHis, complete sequence.
ACCESSION   DQ903887
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5824)
  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 5824)
  AUTHORS   Lamping E, Monk BC, Niimi K, Holmes AR, Tsao S, Kaneko A, Niimi M, 
            Uehara Y, Cannon RD.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-AUG-2006) Oral Sciences, Univ. of Otago, 310 Gt. King 
            St, Dunedin, Otago 9001, New Zealand
REFERENCE   3  (bases 1 to 5824)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5824
                     /organism="Cloning vector pABC3-FlagHis"
                     /mol_type="other DNA"
                     /label=derivative of cloning vector pABC3
                     /note="derivative of cloning vector pABC3"
                     /db_xref="taxon:404020"
     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..3582
                     /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..1909
                     /label=Flag and 6x histidine double affinity tag
                     /note="Flag and 6x histidine double affinity tag"
     CDS             1855..1878
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1879..1896
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      1910..2248
                     /regulatory_class="terminator"
                     /note="Saccharomyces cerevisiae strain AD1-8u- PGK1"
     terminator      1974..2244
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     promoter        2280..2491
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2492..3295
                     /codon_start=1
                     /gene="URA3"
                     /product="orotidine-5-phosphate decarboxylase"
                     /label=URA3
                     /note="OMP decarboxylase"
                     /protein_id="ABI82037.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            2492..3295
                     /gene="URA3"
                     /label=URA3
     misc_feature    3315..3582
                     /label=Saccharomyces cerevisiae PDR5 gene, 3 end
                     /note="Saccharomyces cerevisiae PDR5 gene, 3 end"
     promoter        complement(3636..3654)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3675..3691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3699..3715
                     /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(3723..3753)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3768..3789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4077..4665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4836..5696)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="ABI82038.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5697..5801)
                     /gene="bla"
                     /label=AmpR promoter