LOCUS       Exported                6819 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pY1-gLUC59(GW), complete sequence.
ACCESSION   KY581739
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6819)
  AUTHORS   Bonaldi K, Li Z, Kang SE, Breton G, Pruneda-Paz JL.
  TITLE     Novel surface luciferase reporter for high-throughput quantitative 
            test one-hybrid screens
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6819)
  AUTHORS   Pruneda-Paz JL.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-FEB-2017) Biological Sciences, University of 
            California San Diego, 9500 Gilman Dr, La Jolla, CA 92093, USA
REFERENCE   3  (bases 1 to 6819)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6819
                     /organism="Cloning vector pY1-gLUC59(GW)"
                     /mol_type="other DNA"
                     /db_xref="taxon:2033867"
     misc_recomb     30..154
                     /label=attR1 gateway recombination site
                     /note="attR1 gateway recombination site"
     protein_bind    30..154
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        179..209
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             263..922
                     /codon_start=1
                     /gene="cmR"
                     /product="chloramphenicol acetyltransferase"
                     /label=cmR
                     /note="chloramphenicol resistance protein"
                     /protein_id="ASZ00118.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            263..922
                     /gene="cmR"
                     /label=cmR
     CDS             1264..1569
                     /codon_start=1
                     /gene="ccdB"
                     /product="cell cycle death B cytotoxic protein"
                     /label=ccdB
                     /note="inhibits DNA gyrase"
                     /protein_id="ASZ00119.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            1264..1569
                     /gene="ccdB"
                     /label=ccdB
     misc_recomb     complement(1610..1734)
                     /label=attR2 gateway recombination site
                     /note="attR2 gateway recombination site"
     protein_bind    complement(1610..1734)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1759..1897
                     /regulatory_class="promoter"
                     /note="from pLacZi; CYC1"
     CDS             1898..2824
                     /codon_start=1
                     /gene="gLUC59"
                     /product="cell surface exposed luciferase gLUC59"
                     /label=gLUC59
                     /note="synthetic construct cell surface exposed luciferase 
                     gLUC59 gene (FJ790493)"
                     /protein_id="ASZ00120.1"
                     /translation="MQFSSAIILSAVAGSALATYGSGVKVLFALICIAVAEAKPTENNE
                     DFNIVAVASNFATTDLDADRGKLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCT
                     PKMKKFIPGRCHTYEGDKESAQGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTT
                     GCLKGLANVQCSDLLKKWLPQRCATFASKIQGQVDKNKGAGGDTHGMDELYKANSTVTD
                     IATTVVTITSCEENKCLETEVTTGVTTVTEVETTYTTYCPLPTAKAPVASTSNSTTTPP
                     VSTAEGAAAANAVPAVAAGLLALGAFM"
     gene            1898..2824
                     /gene="gLUC59"
                     /label=gLUC59
     misc_feature    1898..1958
                     /gene="gLUC59"
                     /note="from Candida albicans Pga59/orf19.2767 signal 
                     peptide and first two amino acid residues of mature domain;
                     PGA59"
     misc_feature    1964..2515
                     /gene="gLUC59"
                     /note="from Gaussia princeps luciferase codon-optimized for
                     expression in Candida albicans; gLUC"
     misc_feature    2534..2824
                     /gene="gLUC59"
                     /note="from Candida albicans Pga59/orf19.2767 mature domain
                     and GPI-anchor addition domain; PGA59"
     regulatory      2825..3314
                     /regulatory_class="terminator"
                     /note="from CIp10::ACT1p-gLUC59; PGA59"
     primer_bind     complement(3569..3585)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3593..3609
                     /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(3617..3647)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3662..3683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3971..4559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      4084..4534
                     /label=Col E1ori from pLacZi
                     /note="Col E1ori from pLacZi"
     CDS             complement(4730..5590)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /function="ampicillin resistance"
                     /label=bla
                     /note="from pLacZi"
                     /protein_id="ASZ00121.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4730..5590)
                     /gene="bla"
                     /label=bla
     promoter        complement(5591..5695)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(5800..6603)
                     /codon_start=1
                     /gene="URA3"
                     /product="orotidine-5-phosphate decarboxylase"
                     /label=URA3
                     /note="from pLacZi"
                     /protein_id="ASZ00122.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            complement(5800..6603)
                     /gene="URA3"
                     /label=URA3
     promoter        complement(6604..6819)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter