LOCUS       sequence-402679-        6437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DUGV OTU domain of L protein from a GAL10 promoter with a 
            URA3 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6437)
  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 6437)
  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-DUGV_OTU
FEATURES             Location/Qualifiers
     source          1..6437
                     /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(2425..2441)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2426..2442)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      2444..2691
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2460..2478)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2709..2728)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2710..2728)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2738..2755)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2738..2754)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2747..2769)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2895..3350
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2982..3001)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3192..3213
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3481..4284)
                     /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(4285..4500)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4524..4543)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4643..4665
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4703..4721)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4762..5265
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5302..5406
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5407..5475,5476..6267)
                     /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(5625..5644)
                     /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 aaaagcaggc tacaaaatgg
     1861 acttcttaga cagtctaatt tgggaaaggg ttgttgatga acaatatatc actaacccaa
     1921 cattttgtgt tagtgactat tttgaagtta tcagacagcc aggggatggc aactgttttt
     1981 accatagcat tgcagaactg ttctttgatg tcaaaacacc atctagcttt agaaaagtaa
     2041 aggagcattt acagttagca gcagaagtgt actatgacac tgagccagag gcagtaggaa
     2101 ctggaatctc aaaagatgag tacatcaagg tcgcaatgaa agacaatgaa tggggaggtt
     2161 ccttggaagc ttcaatgtta tcaaagcatc tccaaacaac catcatcttg tgggtggtca
     2221 attcgacaga acaggttacg gccgctatta agtttggacc aggcagggta tcaacagctc
     2281 taaatcttat gcatgttggt cgcacacact tcgatgcgct taggataata gaacagctgg
     2341 agaacaacca acctcaagac aggtaaaacc cagctttctt gtactaagtg gtgatgggct
     2401 gcaggaattc gatatcaagc ttatcgatac cgtcgacctc gagtcatgta attagttatg
     2461 tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga
     2521 caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta
     2581 tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata
     2641 ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cggccggtac
     2701 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc
     2761 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     2821 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     2881 tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2941 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3001 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3061 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3121 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3181 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3241 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3301 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg
     3361 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagggtaata actgatataa
     3421 ttaaattgaa gctctaattt gtgagtttag tatacatgca tttacttata atacagtttt
     3481 ttagttttgc tggccgcatc ttctcaaata tgcttcccag cctgcttttc tgtaacgttc
     3541 accctctacc ttagcatccc ttccctttgc aaatagtcct cttccaacaa taataatgtc
     3601 agatcctgta gagaccacat catccacggt tctatactgt tgacccaatg cgtctccctt
     3661 gtcatctaaa cccacaccgg gtgtcataat caaccaatcg taaccttcat ctcttccacc
     3721 catgtctctt tgagcaataa agccgataac aaaatctttg tcgctcttcg caatgtcaac
     3781 agtaccctta gtatattctc cagtagatag ggagcccttg catgacaatt ctgctaacat
     3841 caaaaggcct ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac cgctaacaat
     3901 acctgggccc accacaccgt gtgcattcgt aatgtctgcc cattctgcta ttctgtatac
     3961 acccgcagag tactgcaatt tgactgtatt accaatgtca gcaaattttc tgtcttcgaa
     4021 gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc ttaactgtgc cctccatgga
     4081 aaaatcagtc aagatatcca catgtgtttt tagtaaacaa attttgggac ctaatgcttc
     4141 aactaactcc agtaattcct tggtggtacg aacatccaat gaagcacaca agtttgtttg
     4201 cttttcgtgc atgatattaa atagcttggc agcaacagga ctaggatgag tagcagcacg
     4261 ttccttatat gtagctttcg acatgattta tcttcgtttc ctgcaggttt ttgttctgtg
     4321 cagttgggtt aagaatactg ggcaatttca tgtttcttca acactacata tgcgtatata
     4381 taccaatcta agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc
     4441 aaaaaaattt caaggaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa
     4501 aaggtggtat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4561 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4621 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4681 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4741 ataataatgg tttcttagac ggatcgcttg cctgtaactt acacgcgcct cgtatctttt
     4801 aatgatggaa taatttggga atttactctg tgtttattta tttttatgtt ttgtatttgg
     4861 attttagaaa gtaaataaag aaggtagaag agttacggaa tgaagaaaaa aaaataaaca
     4921 aaggtttaaa aaatttcaac aaaaagcgta ctttacatat atatttatta gacaagaaaa
     4981 gcagattaaa tagatataca ttcgattaac gataagtaaa atgtaaaatc acaggatttt
     5041 cgtgtgtggt cttctacaca gacaagatga aacaattcgg cattaatacc tgagagcagg
     5101 aagagcaaga taaaaggtag tatttgttgg cgatccccct agagtctttt acatcttcgg
     5161 aaaacaaaaa ctattttttc tttaatttct ttttttactt tctattttta atttatatat
     5221 ttatattaaa aaatttaaat tataattatt tttatagcac gtgatgaaaa ggacccaggt
     5281 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5341 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5401 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5461 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5521 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5581 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5641 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5701 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5761 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5821 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5881 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5941 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6001 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6061 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6121 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6181 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6241 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6301 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6361 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6421 aagatcaaag gatcttc
//
