LOCUS       sequence-402806-        7420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MERS-CoV papain-like protease from a GAL promoter with a 
            URA3 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7420)
  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 7420)
  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: pAG426GAL-MERS-CoV_PLP
FEATURES             Location/Qualifiers
     source          1..7420
                     /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        1054..1495
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1054..1171
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     1466..1489
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     1510..1526
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1510..1526
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    1534..1558
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2522..2546)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2579..2595)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2580..2596)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      2598..2845
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2614..2632)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2863..2882)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2864..2882)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2892..2909)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2892..2908)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2901..2923)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3049..3504
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3136..3155)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3346..3367
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3635..4438)
                     /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(4439..4654)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4678..4697)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4797..4819
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4857..4875)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      4916..6258
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    5248..5295
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        6285..6389
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6390..6458,6459..7250)
                     /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(6608..6627)
                     /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 gtacggatta gaagccgccg agcgggtgac
     1081 agccctccga aggaagactc tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa
     1141 acgcagatgt gcctcgcgcc gcactgctcc gaacaataaa gattctacaa tactagcttt
     1201 tatggttatg aagaggaaaa attggcagta acctggcccc acaaaccttc aaatgaacga
     1261 atcaaattaa caaccatagg atgataatgc gattagtttt ttagccttat ttctggggta
     1321 attaatcagc gaagcgatga tttttgatct attaacagat atataaatgc aaaaactgca
     1381 taaccacttt aactaatact ttcaacattt tcggtttgta ttacttctta ttcaaatgta
     1441 ataaaagtat caacaaaaaa ttgttaatat acctctatac tttaacgtca aggagaaaaa
     1501 accccggatt ctagaactag tggatccccc atcacaagtt tgtacaaaaa agcaggctac
     1561 aaaatgacaa tcgaagtctt agtgactgtc gatggtgtaa attttagaac agtcgttcta
     1621 aataataaga acacttatag atcacagctt ggatgcgttt tctttaatgg tgctgatatt
     1681 tctgacacca ttcctgatga gaaacagaat ggtcacagtt tatatctagc agacaatttg
     1741 actgctgatg aaacaaaggc gcttaaagag ttatatggcc ccgttgatcc tactttctta
     1801 cacagattct attcacttaa ggctgcagtc catgggtgga agatggttgt gtgtgataag
     1861 gtacgttctc tcaaattgag tgataataat tgttatctta atgcagttat tatgacactt
     1921 gatttattga aggacattaa atttgttata cctgctctac agcatgcatt tatgaaacat
     1981 aagggcggtg attcaactga cttcatagcc ctcattatgg cttatggcaa ttgcacattt
     2041 ggtgctccag atgatgcctc tcggttactt cataccgtgc ttgcaaaggc tgagttatgc
     2101 tgttctgcac gcatggtttg gagagagtgg tgcaatgtct gtggcataaa agatgttgtt
     2161 ctacaaggct taaaagcttg ttgttacgtg ggtgtgcaaa ctgttgaaga tctgcgtgct
     2221 cgcatgacat atgtatgcca gtgtggtggt gaacgtcatc ggcaattagt cgaacacacc
     2281 accccctggt tgctgctctc aggcacacca aatgaaaaat tggtgacaac ctccacggcg
     2341 cctgattttg tagcatttaa tgtctttcag ggcattgaaa cggctgttgg ccattatgtt
     2401 catgctcgcc tgaagggtgg tcttatttta aagtttgact ctggcaccgt tagcaagact
     2461 tcagactgga agtgcaaggt gacagatgta cttttccccg gccaaaaata cagtagctaa
     2521 aacccagctt tcttgtacaa agtggtgatg ggctgcagga attcgatatc aagcttatcg
     2581 ataccgtcga cctcgagtca tgtaattagt tatgtcacgc ttacattcac gccctccccc
     2641 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     2701 tttttttata gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt
     2761 tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt
     2821 gggacgctcg aaggctttaa tttgcggccg gtacccaatt cgccctatag tgagtcgtat
     2881 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2941 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     3001 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta
     3061 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3121 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3181 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3241 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3301 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3361 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3421 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3481 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     3541 tatttcacac cgcatagggt aataactgat ataattaaat tgaagctcta atttgtgagt
     3601 ttagtataca tgcatttact tataatacag ttttttagtt ttgctggccg catcttctca
     3661 aatatgcttc ccagcctgct tttctgtaac gttcaccctc taccttagca tcccttccct
     3721 ttgcaaatag tcctcttcca acaataataa tgtcagatcc tgtagagacc acatcatcca
     3781 cggttctata ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca
     3841 taatcaacca atcgtaacct tcatctcttc cacccatgtc tctttgagca ataaagccga
     3901 taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc cttagtatat tctccagtag
     3961 atagggagcc cttgcatgac aattctgcta acatcaaaag gcctctaggt tcctttgtta
     4021 cttcttctgc cgcctgcttc aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat
     4081 tcgtaatgtc tgcccattct gctattctgt atacacccgc agagtactgc aatttgactg
     4141 tattaccaat gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac ttggcggata
     4201 atgcctttag cggcttaact gtgccctcca tggaaaaatc agtcaagata tccacatgtg
     4261 tttttagtaa acaaattttg ggacctaatg cttcaactaa ctccagtaat tccttggtgg
     4321 tacgaacatc caatgaagca cacaagtttg tttgcttttc gtgcatgata ttaaatagct
     4381 tggcagcaac aggactagga tgagtagcag cacgttcctt atatgtagct ttcgacatga
     4441 tttatcttcg tttcctgcag gtttttgttc tgtgcagttg ggttaagaat actgggcaat
     4501 ttcatgtttc ttcaacacta catatgcgta tatataccaa tctaagtctg tgctccttcc
     4561 ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa atttcaagga aaccgaaatc
     4621 aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaaggtg gtatggtgca ctctcagtac
     4681 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     4741 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4801 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     4861 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agtatgatcc
     4921 aatatcaaag gaaatgatag cattgaagga tgagactaat ccaattgagg agtggcagca
     4981 tatagaacag ctaaagggta gtgctgaagg aagcatacga taccccgcat ggaatgggat
     5041 aatatcacag gaggtactag actacctttc atcctacata aatagacgca tataagtacg
     5101 catttaagca taaacacgca ctatgccgtt cttctcatgt atatatatat acaggcaaca
     5161 cgcagatata ggtgcgacgt gaacagtgag ctgtatgtgc gcagctcgcg ttgcattttc
     5221 ggaagcgctc gttttcggaa acgctttgaa gttcctattc cgaagttcct attctctaga
     5281 aagtatagga acttcagagc gcttttgaaa accaaaagcg ctctgaagac gcactttcaa
     5341 aaaaccaaaa acgcaccgga ctgtaacgag ctactaaaat attgcgaata ccgcttccac
     5401 aaacattgct caaaagtatc tctttgctat atatctctgt gctatatccc tatataacct
     5461 acccatccac ctttcgctcc ttgaacttgc atctaaactc gacctctaca ttttttatgt
     5521 ttatctctag tattactctt tagacaaaaa aattgtagta agaactattc atagagtgaa
     5581 tcgaaaacaa tacgaaaatg taaacatttc ctatacgtag tatatagaga caaaatagaa
     5641 gaaaccgttc ataattttct gaccaatgaa gaatcatcaa cgctatcact ttctgttcac
     5701 aaagtatgcg caatccacat cggtatagaa tataatcggg gatgccttta tcttgaaaaa
     5761 atgcacccgc agcttcgcta gtaatcagta aacgcgggaa gtggagtcag gcttttttta
     5821 tggaagagaa aatagacacc aaagtagcct tcttctaacc ttaacggacc tacagtgcaa
     5881 aaagttatca agagactgca ttatagagcg cacaaaggag aaaaaaagta atctaagatg
     5941 ctttgttaga aaaatagcgc tctcgggatg catttttgta gaacaaaaaa gaagtataga
     6001 ttctttgttg gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa atgcagctca
     6061 gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca aaaatgaagc
     6121 acagattctt cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg taaaaatgca
     6181 gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt ctacaaaatg
     6241 aagcacagat gcttcgttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6301 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6361 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6421 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6481 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6541 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     6601 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6661 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6721 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6781 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6841 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6901 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6961 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     7021 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     7081 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     7141 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     7201 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     7261 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     7321 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7381 actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
