LOCUS       sequence-455450-        7418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid enabling yeast-mediated expression of Gaussia luciferase 
            (GLuc).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7418)
  AUTHORS   Rebeck ON, Wallace MJ, Prusa J, Ning J, Evbuomwan EM, Rengarajan S, 
            Habimana-Griffin L, Kwak S, Zahrah D, Tung J, Liao J, Mahmud B, 
            Fishbein SRS, Ramirez Tovar ES, Mehta R, Wang B, Gorelik MG, Helmink
            BA, Dantas G
  TITLE     A yeast-based oral therapeutic delivers immune checkpoint inhibitors
            to reduce intestinal tumor burden.
  JOURNAL   Cell Chem Biol. 2024 Nov 16:S2451-9456(24)00452-5. doi: 
            10.1016/j.chembiol.2024.10.013.
  PUBMED    39571582
REFERENCE   2  (bases 1 to 7418)
  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"; journalName: "Cell Chem 
            Biol. 2024 Nov 16:S2451-9456(24)00452-5. doi: 
            10.1016/j.chembiol.2024.10.013."
COMMENT     Sequence Label: pOGS539_GLuc-ns
FEATURES             Location/Qualifiers
     source          1..7418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(4..592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(84..103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      1091..2433
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(2054..2101)
                     /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)."
     CDS             2693..3487
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2693..3487
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2747..2766)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3357..3376
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      3500..3636
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3781..3852
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     polyA_signal    3874..3922
                     /note="synthetic polyadenylation signal"
     promoter        4105..4518
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             4638..4664
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      5298..5545
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5314..5332)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      5603..5649
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5679..5815
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     promoter        5901..6420
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     misc_feature    7125..7196
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
ORIGIN
        1 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
       61 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      121 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      181 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      241 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      301 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      361 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      421 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      481 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      541 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      601 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      661 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      721 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      781 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc
      841 gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg
      901 ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt
      961 ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga
     1021 gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc
     1081 ggggcgcgcc aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc
     1141 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa
     1201 agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg
     1261 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     1321 gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca
     1381 tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg
     1441 ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag
     1501 gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta
     1561 ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt
     1621 ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt
     1681 cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga
     1741 aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt
     1801 tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc
     1861 aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat
     1921 agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct
     1981 cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt
     2041 tcaaaagcgc tctgaagttc ctatacttcc tagagaatag gaacttcgga ataggaactt
     2101 caaagcgttt ccgaaaacga gcgcttccga aaatgcaacg cgagctgcgc acatacagct
     2161 cactgttcac gtcgcaccta tatctgcgtg ttgcctgtat atatatatac atgagaagaa
     2221 cggcatagtg cgtgtttatg cttaaatgcg tacttatatg cgtctattta tgtaggatga
     2281 aaggtagtct agtacctcct gtgatattat cccattccat gcggggtatc gtatgcttcc
     2341 ttcagcacta ccctttagct gttctatatg ctgccactcc tcaattggat tagtctcatc
     2401 cttcaatgct atcatttcct ttgatattgg atcatacggc gcgcctggtg taccgagaac
     2461 gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag ccagtcggaa
     2521 tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct ggtcacggca
     2581 gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt ataatcgagt
     2641 cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc gcatgattga
     2701 acaagatgga ttgcacgcag gttctccggc ggcttgggtg gagaggctat tcggctatga
     2761 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     2821 gcgtccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     2881 ggcagcgcgg ctatcgtggc tggcgacgac gggcgttcct tgcgcggctg tgctcgacgt
     2941 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3001 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3061 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3121 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     3181 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg tctatgcccg acggcgagga
     3241 tctcgtcgtg acccacggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     3301 ttctggattc atcgactgtg gccgtctggg tgtggcggac cgctatcagg acatagcgtt
     3361 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tccttgtgct
     3421 ttacggtatc gccgcgcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     3481 cttctgaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     3541 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     3601 ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtttaaactc gactctggct
     3661 ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg
     3721 ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga
     3781 catcttggac cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt
     3841 cagctacctc tcaatttcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc
     3901 ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga tctcaatgca
     3961 tactttgtac gttcaaaata caatgcagta gatatattta tgcatattac atataataca
     4021 tatcacatag gaagcaacag gcgcgttgga cttttaattt tcgaggaccg cgaatcctta
     4081 catcacaccc aatcccccac aagtgatccc ccacacacca tagcttcaaa atgtttctac
     4141 tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca
     4201 cccaagcaca gcatactaaa tttcccctct ttcttcctct agggtgtcgt taattacccg
     4261 tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag
     4321 gcaataaaaa tttttatcac gtttcttttt cttgaaaatt tttttttttg atttttttct
     4381 ctttcgatga cctcccattg atatttaagt taataaacgg tcttcaattt ctcaagtttc
     4441 agtttcattt ttcttgttct attacaactt tttttacttc ttgctcatta gaaagaaagc
     4501 atagcaatct aatctaagtt ttaattacaa aagatctttg tcgatcctac catccactcg
     4561 acacacccgc cagcggccgc tgccaagctt ccgagctctc gaattcaaag gaggtaccaa
     4621 ccatgggggg aagttcctat ccctatgacg tgccagacta cgcatccagc ggaggtggat
     4681 caggtaaacc gaccgagaac aatgaagatt tcaacatcgt cgccgtagca tcaaatttcg
     4741 ccacgacgga cttagatgca gacagaggaa aactacctgg gaaaaaactt cccttggaag
     4801 tactgaaaga aatggaggcg aacgcgagaa aagctggctg taccaggggt tgtcttatat
     4861 gtctgtcaca tatcaaatgt actccaaaga tgaagaaatt tattcccggg cgttgccata
     4921 catacgaagg ggataaagag tcagctcaag ggggaatcgg ggaggcgata gttgatatac
     4981 ctgagatccc tggttttaag gacctggagc ccatggagca attcatagcg caggtggatt
     5041 tatgtgtgga ctgcaccacc ggatgtctaa aaggcctagc taacgtacag tgctccgact
     5101 tactaaaaaa atggttaccc cagcgttgtg cgacttttgc ttcaaaaatc cagggccagg
     5161 tggacaaaat aaagggggcg ggaggcgatt gatctagaag ttgtctcctc ctgcactgac
     5221 tgactgatac aatcgatttc tggatccgca ggcctctgct agcttgactg actgagatac
     5281 agcgtacctt cagctcatca tgtaattagt tatgtcacgc ttacattcac gccctccccc
     5341 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     5401 tttttttata gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt
     5461 tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt
     5521 gggacgctcg aaggctttaa tttgcggcca aagcaagtaa aacctctaca aatgtggtat
     5581 tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg
     5641 ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc
     5701 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     5761 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg
     5821 tcagctatcc tgcagggtgt ttaaagatta cggatattta acttacttag aataatgcca
     5881 tttttttgag ttataataat cctacgttag tgtgagcggg atttaaactg tgaggacctt
     5941 aatacattca gacacttctg cggtatcacc ctacttattc ccttcgagat tatatctagg
     6001 aacccatcag gttggtggaa gattacccgt tctaagactt ttcagcttcc tctattgatg
     6061 ttacacctgg acaccccttt tctggcatcc agtttttaat cttcagtggc atgtgagatt
     6121 ctccgaaatt aactaaagca atcacacaat tctctcggat accacctcgg ttgaaactga
     6181 caggtggttt gttacgcatg ctaatgcaaa ggagcctata tacctttggc tcggctgctg
     6241 taacagggaa tataaagggc agcataattt aggagtttag tgaacttgca acatttacta
     6301 ttttcccttc ttacgtaaat atttttcttt ttaattctaa atcaatcttt ttcaattttt
     6361 tgtttgtatt cttttcttgc ttaaatctat aactacaaaa aacacataca taaactaaaa
     6421 atggccaaaa ccttcaacat ctctcagcag gatctggagc tggtggaggt cgccactgag
     6481 aagatcacca tgctctatga ggacaacaag caccatgtcg gggcggccat caggaccaag
     6541 actggggaga tcatctctgc tgtccacatt gaagcctaca ttggcagggt cactgtctgt
     6601 gctgaagcca ttgccattgg gtctgctgtg agcaacgggc agaaggactt tgacaccatt
     6661 gtggctgtca ggcaccccta ctctgatgag gtggacagat ccatcagggt ggtcagcccc
     6721 tgtggcatgt gtagagagct gatctctgac tatgctcctg actgctttgt gctcattgag
     6781 atgaatggca agctggtcaa aaccaccatt gaggaactca tccccctcaa gtacaccagg
     6841 aactaaggtt aatataatta tataaaaata ttatcttctt ttctttatat ctagtgttat
     6901 gtaaaataaa ttgatgacta cggaaagcct ttttatattg tttctttttc attctgagcc
     6961 acttaaattt cgtgaatgtt cttgtaaggg acggtagatt tacaagtgat acaacaaaaa
     7021 gcaaggcgct ttttctaata aaaagaagaa aagcatttaa caattgaaca cctctatatc
     7081 aacgaagaat attcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc
     7141 ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct
     7201 cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga
     7261 tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct
     7321 tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag
     7381 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggc
//
