LOCUS       Exported                7067 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJB036, complete sequence.
ACCESSION   KU904418
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7067)
  AUTHORS   Adeniran A, Stainbrook SC, Bostick JW, Tyo KEJ.
  TITLE     Detection of Peptide Biomarkers by Engineered Yeast Receptors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7067)
  AUTHORS   Pertusi D, Yu J, Adeniran A, Tyo KEJ.
  TITLE     CellSort: A support vector machine algorithm for optimizing and 
            simulating FACS
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 7067)
  AUTHORS   Tyo KEJ., Adeniran A, Stainbrook SC, Bostick JW.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-MAR-2016) Chemical and Biological Engineering, 
            Northwestern University, 2145 Sheridan Rd, Evanston, IL 60208, USA
REFERENCE   4  (bases 1 to 7067)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7067
                     /organism="Cloning vector pJB036"
                     /mol_type="other DNA"
                     /label=derived from Saccharomyces cerevisiae sequence
                     /note="derived from Saccharomyces cerevisiae sequence"
                     /db_xref="taxon:1814217"
     primer_bind     complement(20..36)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      39..286
                     /regulatory_class="terminator"
                     /gene="CYC1"
     gene            39..286
                     /gene="CYC1"
                     /label=CYC1
     terminator      39..286
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     regulatory      complement(303..323)
                     /regulatory_class="promoter"
                     /gene="T7"
                     /note="from bacteriophage T7 RNA polymerase"
     gene            complement(303..323)
                     /gene="T7"
                     /label=T7
     promoter        complement(305..323)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(333..349)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(493..949)
                     /direction=LEFT
                     /label=oriM13
                     /note="oriM13"
     rep_origin      494..949
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     regulatory      889..1078
                     /regulatory_class="promoter"
                     /gene="T3"
     gene            889..1078
                     /gene="T3"
                     /label=T3
     gene            complement(1080..2099)
                     /gene="URA3"
                     /label=URA3
     CDS             complement(1080..1883)
                     /codon_start=1
                     /gene="URA3"
                     /product="URA3"
                     /label=URA3
                     /note="yeast auxotrophic marker; counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /protein_id="AMY56671.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     regulatory      complement(1884..2099)
                     /regulatory_class="promoter"
                     /gene="URA3"
     promoter        complement(1884..2099)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     regulatory      2209..3013
                     /regulatory_class="promoter"
                     /gene="AmpR"
     gene            2209..3013
                     /gene="AmpR"
                     /label=AmpR
     misc_feature    2369..2872
                     /note="Saccharomyces cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        2909..3013
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3014..3874
                     /codon_start=1
                     /product="beta-lactamase"
                     /function="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /label=beta-lactamase
                     /protein_id="AMY56672.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4045..4633
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      4957..4987
                     /regulatory_class="promoter"
                     /gene="lac"
                     /note="from Escherchia coli"
     gene            4957..4987
                     /gene="lac"
                     /label=lac
     promoter        4957..4987
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4995..5011
                     /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     5019..5035
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5056..5074
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      5099..5742
                     /regulatory_class="promoter"
                     /gene="TDH3"
     gene            5099..5742
                     /gene="TDH3"
                     /label=TDH3
     promoter        5099..5742
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase"
     primer_bind     5748..5764
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5766..5771
                     /label=Kozak
                     /note="Kozak"
     CDS             5772..7067
                     /codon_start=1
                     /gene="STE2"
                     /product="alpha-factor pheromone receptor"
                     /label=STE2
                     /note="seven transmembrane-domain GPCR interacts with both 
                     pheromone and a heterotrimeric G protein to initiate the 
                     signaling response that leads to mating between haploid a 
                     and alpha cells; Ste2p"
                     /protein_id="AMY56673.1"
                     /translation="MSDAAPSLSNLFYDPTYNPGQSTINYTSIYGNGSTITFDELQGLV
                     NSTVTQAIMFGVRCGAAALTLIVMWMTSRSRKTPIFIINQVSLFLIILHSALYFKYLLS
                     NYSSVTYALTGFPQFISRGDVHVYGATNIIQVLLVASIETSLVFQIKVIFTGDNFKRIG
                     LMLTSISFTLGIATVTMYFVSAVKGMIVTYNDVSATQDKYFNASTILLASSINFMSFVL
                     VVKLILAIRSRRFLGLKQFDSFHILLIMSCQSLLVPSIIFILAYSLKPNQGTDVLTTVA
                     TLLAVLSLPLSSMWATAANNASKTNTITSDFTTSTDRFYPGTLSSFQTDSINNDAKSSL
                     RSRLYDLYPRRKETTSDKHSERTFVSETADDIEKNQFYQLPTPTSSKNTRIGPFADASY
                     KEGEVEPVDMYTPDTAADEEARKFWTEDNNNL"
     gene            5772..7067
                     /gene="STE2"
                     /label=STE2
     misc_feature    5984..5989
                     /gene="STE2"
                     /label=silent_mutation
                     /note="silent_mutation"
     misc_feature    5988..5993
                     /gene="STE2"
                     /label=silent_mutation
                     /note="silent_mutation"
     misc_feature    6816..6821
                     /gene="STE2"
                     /label=silent_mutation
                     /note="silent_mutation"