LOCUS       sequence-402844-        5953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HIV-1 protease from a GALL promoter with a URA3 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5953)
  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 5953)
  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: pAG416GALL-HIV-1_PR
FEATURES             Location/Qualifiers
     source          1..5953
                     /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     1466..1489
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    1550..1574
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1941..1957)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1942..1958)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      1960..2207
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1976..1994)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2225..2244)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2226..2244)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2254..2271)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2254..2270)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2263..2285)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2411..2866
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2498..2517)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2708..2729
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2997..3800)
                     /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(3801..4016)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4040..4059)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4159..4181
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4219..4237)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4278..4781
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4818..4922
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4923..4991,4992..5783)
                     /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(5141..5160)
                     /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 ggctcgtgac
     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 accccggatc cggaattcca gctgaccacc gactagtgga tcccccatca caagtttgta
     1561 caaaaaagca ggctacaaaa tgccccaaat aacactctgg caaagacccc ttgtcactat
     1621 caagattgga ggacagctca aggaggcgtt gctcgacaca ggggcggacg acactgtttt
     1681 ggaggagatg tctttgccgg gacggtggaa gccgaagatg attggaggga tcggtggctt
     1741 tattaaggtg agacaatacg atcaaatcct gatcgagata tgcggtcaca aggcgattgg
     1801 gacggtactt gtagggccta ctcccgtgaa cataatcggg cgcaaccttc ttacacagat
     1861 tggttgtaca ttgaattttt aaaacccagc tttcttgtac taagtggtga tgggctgcag
     1921 gaattcgata tcaagcttat cgataccgtc gacctcgagt catgtaatta gttatgtcac
     1981 gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac
     2041 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta
     2101 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga
     2161 aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc cggtacccaa
     2221 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     2281 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     2341 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2401 tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2461 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2521 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2581 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2641 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2701 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2761 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2821 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
     2881 ttctccttac gcatctgtgc ggtatttcac accgcatagg gtaataactg atataattaa
     2941 attgaagctc taatttgtga gtttagtata catgcattta cttataatac agttttttag
     3001 ttttgctggc cgcatcttct caaatatgct tcccagcctg cttttctgta acgttcaccc
     3061 tctaccttag catcccttcc ctttgcaaat agtcctcttc caacaataat aatgtcagat
     3121 cctgtagaga ccacatcatc cacggttcta tactgttgac ccaatgcgtc tcccttgtca
     3181 tctaaaccca caccgggtgt cataatcaac caatcgtaac cttcatctct tccacccatg
     3241 tctctttgag caataaagcc gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta
     3301 cccttagtat attctccagt agatagggag cccttgcatg acaattctgc taacatcaaa
     3361 aggcctctag gttcctttgt tacttcttct gccgcctgct tcaaaccgct aacaatacct
     3421 gggcccacca caccgtgtgc attcgtaatg tctgcccatt ctgctattct gtatacaccc
     3481 gcagagtact gcaatttgac tgtattacca atgtcagcaa attttctgtc ttcgaagagt
     3541 aaaaaattgt acttggcgga taatgccttt agcggcttaa ctgtgccctc catggaaaaa
     3601 tcagtcaaga tatccacatg tgtttttagt aaacaaattt tgggacctaa tgcttcaact
     3661 aactccagta attccttggt ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt
     3721 tcgtgcatga tattaaatag cttggcagca acaggactag gatgagtagc agcacgttcc
     3781 ttatatgtag ctttcgacat gatttatctt cgtttcctgc aggtttttgt tctgtgcagt
     3841 tgggttaaga atactgggca atttcatgtt tcttcaacac tacatatgcg tatatatacc
     3901 aatctaagtc tgtgctcctt ccttcgttct tccttctgtt cggagattac cgaatcaaaa
     3961 aaatttcaag gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga attgaaaagg
     4021 tggtatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     4081 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4141 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4201 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4261 taatggtttc ttagacggat cgcttgcctg taacttacac gcgcctcgta tcttttaatg
     4321 atggaataat ttgggaattt actctgtgtt tatttatttt tatgttttgt atttggattt
     4381 tagaaagtaa ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg
     4441 tttaaaaaat ttcaacaaaa agcgtacttt acatatatat ttattagaca agaaaagcag
     4501 attaaataga tatacattcg attaacgata agtaaaatgt aaaatcacag gattttcgtg
     4561 tgtggtcttc tacacagaca agatgaaaca attcggcatt aatacctgag agcaggaaga
     4621 gcaagataaa aggtagtatt tgttggcgat ccccctagag tcttttacat cttcggaaaa
     4681 caaaaactat tttttcttta atttcttttt ttactttcta tttttaattt atatatttat
     4741 attaaaaaat ttaaattata attattttta tagcacgtga tgaaaaggac ccaggtggca
     4801 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4861 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4921 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4981 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5041 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5101 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5161 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5221 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5281 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5341 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5401 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5461 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5521 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5581 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5641 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5701 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5761 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5821 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5881 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5941 tcaaaggatc ttc
//
