LOCUS       sequence-402630-        5843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HIV-2 protease from a GAL-1x promoter with a URA3 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5843)
  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 5843)
  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: pAG416GAL-1x-HIV-2_PR
FEATURES             Location/Qualifiers
     source          1..5843
                     /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"
     primer_bind     1372..1395
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     1416..1432
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1416..1432
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    1440..1464
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1831..1847)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1832..1848)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      1850..2097
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1866..1884)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2115..2134)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2116..2134)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2144..2161)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2144..2160)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2153..2175)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2301..2756
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2388..2407)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2598..2619
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2887..3690)
                     /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(3691..3906)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3930..3949)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4049..4071
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4109..4127)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4168..4671
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4708..4812
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4813..4881,4882..5673)
                     /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(5031..5050)
                     /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 tggagctcgc agatgtgcct cgcgccgcac tgctccgaac
     1081 aataaagatt ctacaatact agcttttatg gttatgaaga ggaaaaattg gcagtaacct
     1141 ggccccacaa accttcaaat gaacgaatca aattaacaac cataggatga taatgcgatt
     1201 agttttttag ccttatttct ggggtaatta atcagcgaag cgatgatttt tgatctatta
     1261 acagatatat aaatgcaaaa actgcataac cactttaact aatactttca acattttcgg
     1321 tttgtattac ttcttattca aatgtaataa aagtatcaac aaaaaattgt taatatacct
     1381 ctatacttta acgtcaagga gaaaaaaccc cggattctag aactagtgga tcccccatca
     1441 caagtttgta caaaaaagca ggctacaaaa tgccgcaatt ctccctgtgg aaaaggcctg
     1501 ttgtgaccgc gtatatcgag gggcaaccag ttgaagttct cctggacact ggggctgatg
     1561 actccatcgt cgcaggtatt gaattgggaa ataactactc ccctaagatt gtggggggca
     1621 taggagggtt catcaacaca aaggagtaca agaatgttga aatcgaggta ttgaacaaaa
     1681 aagtcagagc aaccattatg acaggcgata cgccaatcaa catatttggt cgaaacatct
     1741 tgacggcttt gggtatgtca ctcaatctct aaaacccagc tttcttgtac taagtggtga
     1801 tgggctgcag gaattcgata tcaagcttat cgataccgtc gacctcgagt catgtaatta
     1861 gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga
     1921 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
     1981 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
     2041 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc
     2101 cggtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     2161 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     2221 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2281 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2341 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2401 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2461 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2521 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2581 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2641 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2701 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     2761 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatagg gtaataactg
     2821 atataattaa attgaagctc taatttgtga gtttagtata catgcattta cttataatac
     2881 agttttttag ttttgctggc cgcatcttct caaatatgct tcccagcctg cttttctgta
     2941 acgttcaccc tctaccttag catcccttcc ctttgcaaat agtcctcttc caacaataat
     3001 aatgtcagat cctgtagaga ccacatcatc cacggttcta tactgttgac ccaatgcgtc
     3061 tcccttgtca tctaaaccca caccgggtgt cataatcaac caatcgtaac cttcatctct
     3121 tccacccatg tctctttgag caataaagcc gataacaaaa tctttgtcgc tcttcgcaat
     3181 gtcaacagta cccttagtat attctccagt agatagggag cccttgcatg acaattctgc
     3241 taacatcaaa aggcctctag gttcctttgt tacttcttct gccgcctgct tcaaaccgct
     3301 aacaatacct gggcccacca caccgtgtgc attcgtaatg tctgcccatt ctgctattct
     3361 gtatacaccc gcagagtact gcaatttgac tgtattacca atgtcagcaa attttctgtc
     3421 ttcgaagagt aaaaaattgt acttggcgga taatgccttt agcggcttaa ctgtgccctc
     3481 catggaaaaa tcagtcaaga tatccacatg tgtttttagt aaacaaattt tgggacctaa
     3541 tgcttcaact aactccagta attccttggt ggtacgaaca tccaatgaag cacacaagtt
     3601 tgtttgcttt tcgtgcatga tattaaatag cttggcagca acaggactag gatgagtagc
     3661 agcacgttcc ttatatgtag ctttcgacat gatttatctt cgtttcctgc aggtttttgt
     3721 tctgtgcagt tgggttaaga atactgggca atttcatgtt tcttcaacac tacatatgcg
     3781 tatatatacc aatctaagtc tgtgctcctt ccttcgttct tccttctgtt cggagattac
     3841 cgaatcaaaa aaatttcaag gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga
     3901 attgaaaagg tggtatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3961 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4021 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4081 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4141 gtcatgataa taatggtttc ttagacggat cgcttgcctg taacttacac gcgcctcgta
     4201 tcttttaatg atggaataat ttgggaattt actctgtgtt tatttatttt tatgttttgt
     4261 atttggattt tagaaagtaa ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa
     4321 taaacaaagg tttaaaaaat ttcaacaaaa agcgtacttt acatatatat ttattagaca
     4381 agaaaagcag attaaataga tatacattcg attaacgata agtaaaatgt aaaatcacag
     4441 gattttcgtg tgtggtcttc tacacagaca agatgaaaca attcggcatt aatacctgag
     4501 agcaggaaga gcaagataaa aggtagtatt tgttggcgat ccccctagag tcttttacat
     4561 cttcggaaaa caaaaactat tttttcttta atttcttttt ttactttcta tttttaattt
     4621 atatatttat attaaaaaat ttaaattata attattttta tagcacgtga tgaaaaggac
     4681 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4741 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4801 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4861 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4921 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4981 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5041 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5101 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5161 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5221 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5281 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5341 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5401 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5461 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5521 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5581 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5641 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5701 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5761 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5821 gtagaaaaga tcaaaggatc ttc
//
