LOCUS       sequence-402852-        7174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses protease domain of HHV-3 (VZV) from a GAL promoter with a 
            URA3 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7174)
  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 7174)
  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-HHV-3_PR
FEATURES             Location/Qualifiers
     source          1..7174
                     /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(2276..2300)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2333..2349)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2334..2350)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      2352..2599
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2368..2386)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2617..2636)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2618..2636)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2646..2663)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2646..2662)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2655..2677)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2803..3258
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2890..2909)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3100..3121
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3389..4192)
                     /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(4193..4408)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4432..4451)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4551..4573
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4611..4629)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      4670..6012
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    5002..5049
                     /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        6039..6143
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6144..6212,6213..7004)
                     /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(6362..6381)
                     /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 aaaatggctg ctgaagctga cgaagagaac tgtgaagcgt tatacgtggc tgggtattta
     1621 gccttatatt caaaggacga aggggaatta aatattaccc cagagattgt gcggtccgct
     1681 ttgccgccta ctagtaaaat accaataaac atcgatcatc gaaaagactg tgtcgtgggt
     1741 gaagtaatcg caatcattga ggacatacgc ggaccttttt ttttgggtat cgttagatgc
     1801 cctcaactac atgcggtgct gtttgaagcg gcccattcga atttttttgg aaatagagat
     1861 tctgtattat ctccgctaga acgtgcgtta tacttggtca caaattactt accctccgta
     1921 tccctgtctt caaaacgatt gtccccgaat gagataccag acggtaattt ttttacccat
     1981 gttgcgttat gtgttgttgg aagacgcgtt ggaacagtgg tcaattatga ctgtaccccg
     2041 gaatcttcaa ttgaaccatt ccgggtttta tcgatggaaa gtaaagcgcg gttattgtcg
     2101 ctggttaaag actacgcggg tttaaataaa gtatggaagg ttagcgaaga taaactcgcc
     2161 aaggtgttat tatccacagc cgtgaacaat atgcttttaa gagatagatg ggacgtggtt
     2221 gcaaaacgta gacgcgaagc cggaattatg ggtcacgttt atcttcaggc ttaaaaccca
     2281 gctttcttgt acaaagtggt gatgggctgc aggaattcga tatcaagctt atcgataccg
     2341 tcgacctcga gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc
     2401 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     2461 tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta
     2521 cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg
     2581 ctcgaaggct ttaatttgcg gccggtaccc aattcgccct atagtgagtc gtattacgcg
     2641 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     2701 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     2761 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     2821 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2881 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2941 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3001 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3061 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3121 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3181 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     3241 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3301 acaccgcata gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta
     3361 tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg
     3421 cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa
     3481 atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc
     3541 tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca
     3601 accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa
     3661 aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca gtagataggg
     3721 agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt
     3781 ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa
     3841 tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac
     3901 caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct
     3961 ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca tgtgttttta
     4021 gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg gtggtacgaa
     4081 catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag
     4141 caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc
     4201 ttcgtttcct gcaggttttt gttctgtgca gttgggttaa gaatactggg caatttcatg
     4261 tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt
     4321 cttccttctg ttcggagatt accgaatcaa aaaaatttca aggaaaccga aatcaaaaaa
     4381 aagaataaaa aaaaaatgat gaattgaaaa ggtggtatgg tgcactctca gtacaatctg
     4441 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     4501 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4561 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4621 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagtatg atccaatatc
     4681 aaaggaaatg atagcattga aggatgagac taatccaatt gaggagtggc agcatataga
     4741 acagctaaag ggtagtgctg aaggaagcat acgatacccc gcatggaatg ggataatatc
     4801 acaggaggta ctagactacc tttcatccta cataaataga cgcatataag tacgcattta
     4861 agcataaaca cgcactatgc cgttcttctc atgtatatat atatacaggc aacacgcaga
     4921 tataggtgcg acgtgaacag tgagctgtat gtgcgcagct cgcgttgcat tttcggaagc
     4981 gctcgttttc ggaaacgctt tgaagttcct attccgaagt tcctattctc tagaaagtat
     5041 aggaacttca gagcgctttt gaaaaccaaa agcgctctga agacgcactt tcaaaaaacc
     5101 aaaaacgcac cggactgtaa cgagctacta aaatattgcg aataccgctt ccacaaacat
     5161 tgctcaaaag tatctctttg ctatatatct ctgtgctata tccctatata acctacccat
     5221 ccacctttcg ctccttgaac ttgcatctaa actcgacctc tacatttttt atgtttatct
     5281 ctagtattac tctttagaca aaaaaattgt agtaagaact attcatagag tgaatcgaaa
     5341 acaatacgaa aatgtaaaca tttcctatac gtagtatata gagacaaaat agaagaaacc
     5401 gttcataatt ttctgaccaa tgaagaatca tcaacgctat cactttctgt tcacaaagta
     5461 tgcgcaatcc acatcggtat agaatataat cggggatgcc tttatcttga aaaaatgcac
     5521 ccgcagcttc gctagtaatc agtaaacgcg ggaagtggag tcaggctttt tttatggaag
     5581 agaaaataga caccaaagta gccttcttct aaccttaacg gacctacagt gcaaaaagtt
     5641 atcaagagac tgcattatag agcgcacaaa ggagaaaaaa agtaatctaa gatgctttgt
     5701 tagaaaaata gcgctctcgg gatgcatttt tgtagaacaa aaaagaagta tagattcttt
     5761 gttggtaaaa tagcgctctc gcgttgcatt tctgttctgt aaaaatgcag ctcagattct
     5821 ttgtttgaaa aattagcgct ctcgcgttgc atttttgttt tacaaaaatg aagcacagat
     5881 tcttcgttgg taaaatagcg ctttcgcgtt gcatttctgt tctgtaaaaa tgcagctcag
     5941 attctttgtt tgaaaaatta gcgctctcgc gttgcatttt tgttctacaa aatgaagcac
     6001 agatgcttcg ttcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     6061 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     6121 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     6181 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6241 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6301 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     6361 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     6421 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6481 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6541 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6601 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6661 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6721 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6781 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6841 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6901 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6961 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     7021 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     7081 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7141 cgtcagaccc cgtagaaaag atcaaaggat cttc
//
