LOCUS       Exported                7658 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Integrative expression vector pCYC1lac, complete sequence.
ACCESSION   KF366486
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7658)
  AUTHORS   Vickers CE, Bydder SF, Nielsen LK.
  TITLE     Yeast expression vectors with dominant markers and dual gene 
            expression cassettes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7658)
  AUTHORS   Bydder SF, Vickers CE.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JUL-2013) AIBN, The University of Queensland, Crn 
            College and Cooper Roads (Bldg 75), St Lucia, QLD 4072, Australia
REFERENCE   3  (bases 1 to 7658)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7658
                     /organism="Integrative expression vector pCYC1lac"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
                     /db_xref="taxon:1412981"
     CDS             18..3092
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /protein_id="AGZ03683.1"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYG
                     QDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQ
                     ISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIID
                     ERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIEN
                     GLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLW
                     YTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWS
                     LGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVP
                     KWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSL
                     IKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQ
                     TIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQ
                     LWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELG
                     NKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWK
                     AAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDV
                     EVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPY
                     VFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDG
                     FHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     gene            18..3092
                     /gene="lacZ"
                     /label=lacZ
     primer_bind     complement(38..54)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3093..3340
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     regulatory      3093..3328
                     /regulatory_class="terminator"
                     /note="CYC1 terminator from Saccharomyces cerevisiae"
     CDS             complement(3804..4607)
                     /codon_start=1
                     /gene="URA3"
                     /product="orotidine-5-phosphate (OMP) decarboxylase"
                     /label=URA3
                     /protein_id="AGZ03684.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            complement(3804..4607)
                     /gene="URA3"
                     /label=URA3
     promoter        complement(4608..4823)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     promoter        5113..5217
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5218..6078
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="confers ampicillin resistance in Escherichia coli"
                     /protein_id="AGZ03685.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            5218..6078
                     /gene="bla"
                     /label=bla
     rep_origin      6249..6837
                     /label=prokaryotic pMB1 ori
                     /note="prokaryotic pMB1 ori"
     protein_bind    7125..7146
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      7128..7217
                     /regulatory_class="promoter"
                     /note="beta-galactosidase promoter region"
     misc_feature    7128..7143
                     /label=CAP binding site
                     /note="CAP binding site"
     promoter        7161..7191
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_feature    7161..7190
                     /label=RNA polymerase binding site
                     /note="RNA polymerase binding site"
     misc_feature    7197..7217
                     /label=lac repressor binding site
                     /note="lac repressor binding site"
     protein_bind    7199..7215
                     /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)."
     primer_bind     7223..7239
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7260..7278
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     7307..7323
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      7366..7656
                     /regulatory_class="promoter"
                     /note="CYC1 promoter from Saccharomyces cerevisiae"