LOCUS       sequence-402869-        7550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human YES1 kinase from a GAL10 promoter with a URA3 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7550)
  AUTHORS   Hong SJ, Resnick SJ, Iketani S, Cha JW, Albert BA, Fazekas CT, Chang
            CW, Liu H, Dagan S, Abagyan MR, Fajtova P, Culbertson B, Brace B, 
            Reddem ER, Forouhar F, Glickman JF, Balkovec JM, Stockwell BR, 
            Shapiro L, O'Donoghue AJ, Sabo Y, Freundlich JS, Ho DD, Chavez A
  TITLE     A multiplex method for rapidly identifying viral protease 
            inhibitors.
  JOURNAL   Mol Syst Biol. 2025 Feb;21(2):158-172. doi: 
            10.1038/s44320-024-00082-1. Epub 2025 Jan 6.
  PUBMED    39762652
REFERENCE   2  (bases 1 to 7550)
  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"; doi: 
            "10.1038/s44320-024-00082-1"; journalName: "Mol Syst Biol"; date: 
            "2025-02"; volume: "21"; issue: "2"; pages: "158-172"
COMMENT     Sequence Label: pAG416GAL10-YES1-6stop
FEATURES             Location/Qualifiers
     source          1..7550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1108..1549)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(1114..1137)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    1432..1549
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     protein_bind    1827..1851
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3538..3554)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3539..3555)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      3557..3804
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3573..3591)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3822..3841)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3823..3841)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3851..3868)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3851..3867)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3860..3882)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4008..4463
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4095..4114)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4305..4326
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4594..5397)
                     /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(5398..5613)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5637..5656)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5756..5778
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5816..5834)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5875..6378
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6415..6519
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6520..6588,6589..7380)
                     /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     complement(6738..6757)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctcta tgaccatgat tacgccaagc ggggtgtgtt
     1081 gttggtgggt tgggtgtata gttttttctc cttgacgtta aagtatagag gtatattaac
     1141 aattttttgt tgatactttt atgacatttg aataagaagt aatacaaact gaaaatgttg
     1201 aaagtattag ttaaagtggt tatgcagctt ttccatttat atatctgtta atagatcaaa
     1261 aatcatcgct tcgctgatta attaccccag aaataaggct aaaaaactaa tcgcattatc
     1321 atcctatggt tgttaatttg attcgttaat ttgaaggttt gtggggccag gttactgcca
     1381 atttttcctc ttcataacca taaaagctag tattgtagaa tctttattgt tcggagcagt
     1441 gcggcgcgag gcacatctgc gtttcaggaa cgcgaccggt gaagacgagg acgcacggag
     1501 gagagtcttc cgtcggaggg ctgtcgcccg ctcggcggct tctaatccgt acttcaatat
     1561 agcaatgagc agttaagcgt attactgaaa gttccaaaga gaaggttttt ttaggctaag
     1621 ataatggggc tctttacatt tccacaacat ataagtaaga ttagatatgg atatgtatat
     1681 ggtggtaatg ccatgtaata tgattattaa acttctttgc gtccatccaa aaaaaaagta
     1741 agaatttttg aaaattcaat ataaatgtct aaaggtgaag aattattcag aattccagct
     1801 gaccaccgac tagtggatcc cccatcacaa gtttgtacaa aaaagttggc atgggctgca
     1861 ttaaaagtaa agaaaacaaa agtccagcca ttaaatacag acctgaaaat actccagagc
     1921 ctgtcagtac aagtgtgagc cattatggag cagaacccac tacagtgtca ccatgtccgt
     1981 catcttcagc aaagggaaca gcagttaatt tcagcagtct ttccatgaca ccatttggag
     2041 gatcctcagg ggtaacgcct tttggaggtg catcttcctc attttcagtg gtgccaagtt
     2101 catatcctgc tggtttaaca ggtggtgtta ctatatttgt ggccttatat gattatgaag
     2161 ctagaactac agaagacctt tcatttaaga agggtgaaag atttcaaata attaacaata
     2221 cggaaggaga ttggtgggaa gcaagatcaa tcgctacagg aaagaatggt tatatcccga
     2281 gcaattatgt agcgcctgca gattccattc aggcagaaga atggtatttt ggcaaaatgg
     2341 ggagaaaaga tgctgaaaga ttacttttga atcctggaaa tcaacgaggt attttcttag
     2401 taagagagag tgaaacaact aaaggtgctt attccctttc tattcgtgat tgggatgaga
     2461 taaggggtga caatgtgaaa cactacaaaa ttaggaaact tgacaatggt ggatactata
     2521 tcacaaccag agcacaattt gatactctgc agaaattggt gaaacactac acagaacatg
     2581 ctgatggttt atgccacaag ttgacaactg tgtgtccaac tgtgaaacct cagactcaag
     2641 gtctagcaaa agatgcttgg gaaatccctc gagaatcttt gcgactagag gttaaactag
     2701 gacaaggatg tttcggcgaa gtgtggatgg gaacatggaa tggaaccacg aaagtagcaa
     2761 tcaaaacact aaaaccaggt acaatgatgc cagaagcttt ccttcaagaa gctcagataa
     2821 tgaaaaaatt aagacatgat aaacttgttc cactatatgc tgttgtttct gaagaaccaa
     2881 tttacattgt cactgaattt atgtcaaaag gaagcttatt agatttcctt aaggaaggag
     2941 atggaaagta tttgaagctt ccacagctgg ttgatatggc tgctcagatt gctgatggta
     3001 tggcatatat tgaaagaatg aactatattc accgagatct tcgggctgct aatattcttg
     3061 taggagaaaa tcttgtgtgc aaaatagcag actttggttt agcaaggtta attgaagaca
     3121 atgaatacac agcaagacaa ggtgcaaaat ttccaatcaa atggacagct cctgaagctg
     3181 cactgtatgg tcggtttaca ataaagtctg atgtctggtc atttggaatt ctgcaaacag
     3241 aactagtaac aaagggccga gtgccatatc caggtatggt gaaccgtgaa gtactagaac
     3301 aagtggagcg aggatacagg atgccgtgcc ctcagggctg tccagaatcc ctccatgaat
     3361 tgatgaatct gtgttggaag aaggaccctg atgaaagacc aacatttgaa tatattcagt
     3421 ccttcttgga agactacttc actgctacag agccacagta ccagccagga gaaaatttat
     3481 acccaacttt cttgtactaa gtggtgatgg gctgcaggaa ttcgatatca agcttatcga
     3541 taccgtcgac ctcgagtcat gtaattagtt atgtcacgct tacattcacg ccctcccccc
     3601 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     3661 ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
     3721 ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg
     3781 ggacgctcga aggctttaat ttgcggccgg tacccaattc gccctatagt gagtcgtatt
     3841 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     3901 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     3961 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag
     4021 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4081 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4141 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4201 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4261 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4321 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4381 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4441 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     4501 atttcacacc gcatagggta ataactgata taattaaatt gaagctctaa tttgtgagtt
     4561 tagtatacat gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa
     4621 atatgcttcc cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt
     4681 tgcaaatagt cctcttccaa caataataat gtcagatcct gtagagacca catcatccac
     4741 ggttctatac tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat
     4801 aatcaaccaa tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat
     4861 aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga
     4921 tagggagccc ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac
     4981 ttcttctgcc gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt
     5041 cgtaatgtct gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt
     5101 attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa
     5161 tgcctttagc ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt
     5221 ttttagtaaa caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt
     5281 acgaacatcc aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt
     5341 ggcagcaaca ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat
     5401 ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg gttaagaata ctgggcaatt
     5461 tcatgtttct tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct
     5521 tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa accgaaatca
     5581 aaaaaaagaa taaaaaaaaa atgatgaatt gaaaaggtgg tatggtgcac tctcagtaca
     5641 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     5701 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     5761 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     5821 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacggatcgc
     5881 ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg gaataatttg ggaatttact
     5941 ctgtgtttat ttatttttat gttttgtatt tggattttag aaagtaaata aagaaggtag
     6001 aagagttacg gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc
     6061 gtactttaca tatatattta ttagacaaga aaagcagatt aaatagatat acattcgatt
     6121 aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac acagacaaga
     6181 tgaaacaatt cggcattaat acctgagagc aggaagagca agataaaagg tagtatttgt
     6241 tggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt ttctttaatt
     6301 tcttttttta ctttctattt ttaatttata tatttatatt aaaaaattta aattataatt
     6361 atttttatag cacgtgatga aaaggaccca ggtggcactt ttcggggaaa tgtgcgcgga
     6421 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6481 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     6541 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     6601 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     6661 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6721 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6781 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6841 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6901 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6961 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     7021 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     7081 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     7141 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     7201 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     7261 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7321 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     7381 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     7441 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7501 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
