LOCUS       sequence-267643-        6806 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mutant yeast STE2 coding sequence in vector for expression in yeast 
            S. cerevisiae, with Gal1 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6806)
  TITLE     Wagner Lab plasmids
REFERENCE   2  (bases 1 to 6806)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pYES2-STE2-1D4
FEATURES             Location/Qualifiers
     source          1..6806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..442
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1..118
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     413..436
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     474..493
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        474..492
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1554..1801
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1570..1588)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1878..1895)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2049..2637)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2129..2148)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2808..3599,3600..3668))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3431..3450
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(3764..4567)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4568..4783)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5384..6261
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6272..6785)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6467..6488)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6679..6698
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cggattagaa gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc
       61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
      121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
      181 tggccccaca aaccttcaaa tgaacgaatc aaattaacaa ccataggatg ataatgcgat
      241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
      301 aacagatata taaatgcaaa aactgcataa ccactttaac taatactttc aacattttcg
      361 gtttgtatta cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc
      421 tctatacttt aacgtcaagg agaaaaaacc ccggatcgga ctactagcag ctgtaatacg
      481 actcactata gggaatatta agcttggtac cgagctcgga tccactagta acggccgcca
      541 gtgtgctgga attcataatg tctgatgcgg ctccttcatt gagcaatcta ttttatgatc
      601 caacgtataa tcctggtcaa agcaccatta actacacttc catatatggg aatggatcta
      661 ccatcacttt cgatgagttg caaggtttag ttaacagtac tgttactcag gccattatgt
      721 ttggtgtcag atcaggcgcc gctgctttga ctttgattgt catgtggatg acatcgagaa
      781 gcagaaaaac gccgattttc attatcaacc aagtttcatt gtttttaatc attttgcatt
      841 ctgcactcta ttttaaatat ttactgtcta attactcttc agtgacttac gctctcaccg
      901 gatttcctca gttcatcagt agaggtgacg ttcatgttta tggtgctaca aatataattc
      961 aagtccttct tgtggcttct attgagactt cactggtgtt tcagataaaa gttattttca
     1021 caggcgacaa cttcaaaagg ataggtttga tgctgacgtc gatatctttc actttaggga
     1081 ttgctacagt taccatgtat tttgtaagcg ctgttaaagg tatgattgtg acttataatg
     1141 atgttagtgc cacccaagat aaatacttca atgcatccac aattttactt gcatcctcaa
     1201 taaactttat gtcatttgtc ctggtagtta aattgatttt agctattaga tcaagaagat
     1261 tccttggtct caagcagttc gatagtttcc atattttact cataatgtca gctcaatctt
     1321 tgttggtacc atcgataata ttcatcctcg catacagttt gaaaccaaac cagggaacag
     1381 atgtcttgac tactgttgca acattacttg ctgtattgtc tttaccatta tcatcaatgt
     1441 gggccacggc tgctaataat gcgtcgacca ccgtctccaa gacagagacc agccaagtgg
     1501 cgcctgccta aggatcctaa gcggccgctc gagcatgcat ctagagggcc gcatcatgta
     1561 attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga
     1621 aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt
     1681 aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg
     1741 taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg
     1801 caagctgcgg ccctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1861 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1921 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1981 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2041 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2101 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2161 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2221 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2281 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2341 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2401 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2461 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2521 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2581 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2641 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2701 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2761 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2821 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2881 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2941 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3001 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3061 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3121 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3181 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3241 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3301 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3361 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3421 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3481 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3541 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3601 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3661 atactcatac tcttcctttt tcaatgggta ataactgata taattaaatt gaagctctaa
     3721 tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt tgctggccgc
     3781 atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct accttagcat
     3841 cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct gtagagacca
     3901 catcatccac ggttctatac tgttgaccca atgcgtctcc cttgtcatct aaacccacac
     3961 cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct ctttgagcaa
     4021 taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt
     4081 ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg cctctaggtt
     4141 cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg cccaccacac
     4201 cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca gagtactgca
     4261 atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact
     4321 tggcggataa tgcctttagc ggcttaactg tgccctccat ggaaaaatca gtcaagatat
     4381 ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac tccagtaatt
     4441 ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg tgcatgatat
     4501 taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta tatgtagctt
     4561 tcgacatgat ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg gttaagaata
     4621 ctgggcaatt tcatgtttct tcaacactac atatgcgtat atataccaat ctaagtctgt
     4681 gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa
     4741 accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaaaagctag cttatcgatg
     4801 ataagctgtc aaacatgaga attaattcca cggactatag actataccta gtatactccg
     4861 tctactgtac gatacacttc cgctcaggtc cttgtccttt aacgaggcct taccactctt
     4921 ttgttactct attgatccag ctcagcaaag gcagtgtgat ctaagattct atcttcgcga
     4981 tgtagtaaaa ctagctagac cgagaaagag actagaaatg caaaaggcac ttctacaatg
     5041 gctgccatca ttattatccg atgtgacgct gcagcttctc aatgatattc gaatacgctt
     5101 tgaggagata cagcctaata tccgacaaac tgttttacag atttacgatc gtacttgtta
     5161 cccatcattg aattttgaac atccgaacct gggagttttc cctgaaacag atagtatatt
     5221 tgaacctgta taataatata tagtctagcg ctttacggaa gacaatgtat gtatttcggt
     5281 tcctggagaa actattgcat ctattgcata ggtaatcttg cacgtcgcat ccccggttca
     5341 ttttctgcgt ttccatcttg cacttcaata gcatatcttt gttaacgaag catctgtgct
     5401 tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     5461 ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg
     5521 tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     5581 ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga
     5641 atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa
     5701 gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt
     5761 tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc
     5821 ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca
     5881 ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt
     5941 tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt
     6001 cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg
     6061 attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata
     6121 aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg
     6181 taggttatat agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt
     6241 tgtggaagcg gtattcgcaa tgggaagctc caccccggtt gataatcaga aaagccccaa
     6301 aaacaggaag attgtataag caaatattta aattgtaagc gttaatattt tgttaaaatt
     6361 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     6421 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     6481 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     6541 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     6601 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     6661 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     6721 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     6781 cgcgtgggga tcgatccact agtcca
//
