LOCUS       sequence-311078-        9141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYES2 derivative reporter plasmid encoding PREV1-Saccharomyces 
            cerevisiae FEX1-MFaT; as well as PGAL1-Homo sapiens 
            ADORA2A-EGFP-10HIS-MFaT..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9141)
  AUTHORS   Yoo JI, Seppala S, O'Malley MA
  TITLE     Engineered fluoride sensitivity enables biocontainment and selection
            of genetically-modified yeasts.
  JOURNAL   Nat Commun. 2020 Oct 29;11(1):5459. doi: 10.1038/s41467-020-19271-1.
  PUBMED    33122649
REFERENCE   2  (bases 1 to 9141)
  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: "Nat 
            Commun."; date: "2020-10-29"; volume: "11(1):5459. doi"; pages: " 
            10.1038/s41467-020-19271-1"
COMMENT     Sequence Label: pEFEX3-A2aGH
FEATURES             Location/Qualifiers
     source          1..9141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(100..891,892..960))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     RBS             2080..2088
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     rep_origin      3857..4737
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(4806..5261)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     5155..5174
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5280..5722
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    5280..5397
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     5693..5716
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     5754..5773
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5754..5772
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(7165..7167,7168..7170,7171..7881)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7165..7167,7168..7170,7171..7881)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7210..7231)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7471..7490)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7818..7839
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7894..7923
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     terminator      7994..8241
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(8010..8028)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(8311..8328)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8482..9070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8562..8581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
       61 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      121 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      181 gtagataact acgatacggg agcgcttacc atctggcccc agtgctgcaa tgataccgcg
      241 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      301 gcgcagaagt ggtcctgcaa ctttatccgc ctccattcag tctattaatt gttgccggga
      361 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttggca ttgctacagg
      421 catcgtggtg tcactctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
      481 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
      541 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
      601 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
      661 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
      721 ggataatagt gtatcacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
      781 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
      841 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
      901 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
      961 actctactcc tcgtcaattc tcttaggatt cgattcacat tcatcttttt ttagctacta
     1021 ataggataaa ttataggaat ttataacaaa ttgaaatatg gcaggcagca aaattaaaat
     1081 tgtcttagtt ttttagtgta tagaagtgaa tagctatata aagtatgtgt aaagttggta
     1141 acggaacgaa aaatagaaaa ggatattaca tgggaaaaca tgttgtttac ggagaaatga
     1201 aaagtatatt gtattttgta cgagctaaaa gtacagtggg aacaaagtcg attttgttac
     1261 atctacactg ttgttatcag atctcgagct attacaagtc ttcttcagaa ataagctttt
     1321 gttatcagga gacggtgact gaggttcctt gaccccagta gtccatacca tagtaagaac
     1381 ccgtacagta atagataccc atgtcttcaa ctcttaagct aacaaatcgg gttggtcaat
     1441 cctctagtcc aagcatagga tccgagagtt atcaccaata agcaatatga aattgcaatc
     1501 gaaacagtat aataaataag catgttaata aatgatagct tatacccttc attgataaac
     1561 gtgctaatgg tactcaaagt gccgcaaaat ccggatatta acgcagaaac aatatgacat
     1621 gaattcaacg agttcacaat tggaacgtcc gtagaaaagt gttttttacc acgttgcacc
     1681 atggtaaata tgccaatcaa taaagtggcg aaaacgtttg ccaagaaagt acccaatgga
     1741 aactttttgt ttgttttatt aaacatttct gccaaccaat atcttaggaa accagcgaaa
     1801 attccaaata gacatggtag tgtccacttg cccctcgaat agttctcata atacgcacac
     1861 agcaccacaa acaaaataat taatggaaaa gctagtgcat acgcgagttt atccacaata
     1921 tcaaaaaatt tcttaaaaaa tggtgctgga gttttgaatt taaactctag aatatttttt
     1981 tccatttctt gtgtatcaac atcaccagca ttttccttta cagtgtctga gggaggggta
     2041 tatggcttcg aaaacgaact agatccataa gcaacaataa cctccttacc aagttgccta
     2101 ccgaaaataa gactacccat ggaaaccata agatgaacga gcaagacaga taaaaactcc
     2161 attataccgt aagctctgtt cggcagtttt gtgtggttag cgatattacc gttagttaag
     2221 tttgtagagt gttcgaacat ctctagcagc atactagaaa aggaagacag ggcaccacag
     2281 tacccagtag ttacacctaa aaacaagact tggtgatctt tcatccaagt gtaagcattt
     2341 aaagattgca taatccccat tagcatgcag gatgaacaat tagaccagag aacggtcgga
     2401 gcagtaacga aggccggagt gtaggtggaa agcgccgtta tagcctgacg tgtttcagta
     2461 cctaatatgc agaacgtggt gaatgtacaa aaaacatgaa tataatggct caacttgtga
     2521 tttgatatga ccggattgaa aatcattttg aattctcgct ggatatgcct agaaatgcag
     2581 tatcgattta ttttgagaaa atctgtcaac tcgttacttc tttgataggg aaatctttag
     2641 ttaaatttgt ttaaaattaa gaaccaggtc tttagactat gccaaaaggt attggtgctt
     2701 aaaggttgcc gtaaactcta actctatata tctctaagcc aatgtttaag ttgatgcgat
     2761 gtgcatgatc acttcacatt tttttcatga ttttgttttc ttccttagct acttttttcc
     2821 gtcttggtct gtgtaaaaaa aaaatcgctt cactgaagaa aaatataaat aaaaaaagga
     2881 catcagcaag ttcgtagtca ataaatgtga tgagtaattt gcgataatct accccaactc
     2941 aaagcattca attcaaatga aagaagagtt cgtgaatttt aatctggctt acagattgga
     3001 tcatagcttc taatatatct tcctttttat tttaaagtga ttaaaatatg tggatgtctg
     3061 cattaaggtg aaggcagact atagtgaaaa atagtagcaa aggcttcata taatgaacta
     3121 cctagttgta ttcaattgga atctgtgaac gcggcattaa aagaatattt gagctttttt
     3181 tgattgtatt aacctgaagc tatcacggtt tagactcatt aaaagaaaaa tatccggttg
     3241 tatcggataa caacacaggt cagctcttat cgatgataag ctgtcaaaga tgagaattaa
     3301 ttccacggac tatagactat actagatact ccgtctactg tacgatacac ttccgctcag
     3361 gtccttgtcc tttaacgagg ccttaccact cttttgttac tctattgatc cagctcagca
     3421 aaggcagtgt gatctaagat tctatcttcg cgatgtagta aaactagcta gaccgagaaa
     3481 gagactagaa atgcaaaagg cacttctaca atggctgcca tcattattat ccgatgtgac
     3541 gctgcagctt ctcaatgata ttcgaatacg ctttgaggag atacagccta atatccgaca
     3601 aactgtttta cagatttacg atcgtacttg ttacccatca ttgaattttg aacatccgaa
     3661 cctgggagtt ttccctgaaa cagatagtat atttgaacct gtataataat atatagtcta
     3721 gcgctttacg gaagacaatg tatgtatttc ggttcctgga gaaactattg catctattgc
     3781 ataggtaatc ttgcacgtcg catccccggt tcattttctg cgtttccatc ttgcacttca
     3841 atagcatatc tttgttaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
     3901 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     3961 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
     4021 aacgcgacga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     4081 aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac
     4141 aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc
     4201 ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt
     4261 tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc
     4321 ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc
     4381 gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg
     4441 atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta
     4501 cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac
     4561 tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag
     4621 tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca
     4681 gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatggg
     4741 aagctccacc ccggttgata atcagaaaag ccccaaaaac aggaagattg tataagcaaa
     4801 tatttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     4861 ctcatttttt aacgaatagc ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     4921 cgagataggg ttgagtgttg ttccagtttc caacaagagt ccactattaa agaacgtgga
     4981 ctccaacgtc aaagggcgaa aaagggtcta tcagggcgat ggcccactac gtgaaccatc
     5041 accctaatca agttttttgg ggtcgaggtg ccgtaaagca gtaaatcgga agggtaaacg
     5101 gatgccccca tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     5161 gaaagcgaaa ggagcggggg ctagggcggt gggaagtgta ggggtcacgc tgggcgtaac
     5221 caccacaccc gccgcgctta atggggcgct acagggcgcg tggggatgat ccactagtac
     5281 ggattagaag ccgccgagcg ggtgacagcc ctccgaagga agactctcct ccgtgcgtcc
     5341 tcgtcctcac cggtcgcgtt cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac
     5401 aataaagatt ctacaatact agcttttatg gttatgaaga ggaaaaattg gcagtaacct
     5461 ggccccacaa accttcaaat gaacgaatca aattaacaac cataggatga taatgcgatt
     5521 agttttttag ccttatttct ggggtaatta atcagcgaag cgatgatttt tgatctatta
     5581 acagatatat aaatgcaaaa actgcattaa ccactttaac taatactttc aacattttcg
     5641 gtttgtatta cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc
     5701 tctatacttt aacgtcaagg agaaaaaacc ccggatcgga ctactagcag ctgtaatacg
     5761 actcactata gggaatatta agcttggatc atgaaggttt tgattgtctt gttggctatc
     5821 ttcgctgctt tgcccttggc tctagctcaa ccggttattt ctactaccgt cggttccgct
     5881 gcagaaggct ctttggacaa gagagaagct cggccggacg tcatgcccat catgggctcc
     5941 tcggtgtaca tcacggtgga gctggccatt gctgtgctgg ccatcctggg caatgtgctg
     6001 gtgtgctggg ccgtgtggct caacagcaac ctgcagaacg tcaccaacta ctttgtggtg
     6061 tcactggcgg cggccgacat cgcagtgggt gtgctcgcca tcccctttgc catcaccatc
     6121 agcaccgggt tctgcgctgc ctgccacggc tgcctcttca ttgcctgctt cgtcctggtc
     6181 ctcacgcaga gctccatctt cagtctcctg gccatcgcca ttgaccgcta cattgccatc
     6241 cgcatcccgc tccggtacaa tggcttggtg accggcacga gggctaaggg catcattgcc
     6301 atctgctggg tgctgtcgtt tgccatcggc ctgactccca tgctaggttg gaacaactgc
     6361 ggtcagccaa aggagggcaa gaaccactcc cagggctgcg gggagggcca agtggcctgt
     6421 ctctttgagg atgtggtccc catgaactac atggtgtact tcaacttctt tgcctgtgtg
     6481 ctggtgcccc tgctgctcat gctgggtgtc tatttgcgga tcttcctggc ggcgcgacga
     6541 cagctgaagc agatggagag ccagcctctg ccgggggagc gggcacggtc cacactgcag
     6601 aaggaggtcc atgctgccaa gtcactggcc atcattgtgg ggctctttgc cctctgctgg
     6661 ctgcccctac acatcatcaa ctgcttcact ttcttctgcc ccgactgcag ccacgcccct
     6721 ctctggctca tgtacctggc catcgtcctc tcccacacca attcggttgt gaatcccttc
     6781 atctacgcct accgtatccg cgagttccgc cagaccttcc gcaagatcat tcgcagccac
     6841 gtcctgaggc agcaagaacc tttcaaggca gctggcacca gtgcccgggt cttggcagct
     6901 catggcagtg acggagagca ggtcagcctc cgtctcaacg gccacccgcc aggagtgtgg
     6961 gccaacggca gtgctcccca ccctgagcgg aggcccaatg gctacgccct ggggctggtg
     7021 agtggaggga gtgcccaaga gtcccagggg aacacgggcc tcccagacgt ggagctcctt
     7081 agccatgagc tcaagggagt gtgcccagag ccccctggcc tagatgaccc cctggcccag
     7141 gatggagcag gagtgtcccc gcggatggtg agcaagggcg aggagctgtt caccggggtg
     7201 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     7261 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     7321 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     7381 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     7441 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     7501 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     7561 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     7621 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     7681 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     7741 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     7801 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     7861 ggcatggacg agctgtacaa gcacgtgtca tcacatcatc atcatcatca tcatcatcat
     7921 cattaataac ttaagagttg aagacatggg tatctattac tgtacggctc gagcatgcat
     7981 ctagagggcc gcatcatgta attagttatg tcacgcttac attcacgccc tccccccaca
     8041 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     8101 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     8161 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
     8221 cgctcgaagg ctttaatttg cggccctgca ttaatgaatc ggccaacgcg cggggagagg
     8281 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     8341 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     8401 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaagccca ggaaccgtaa
     8461 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     8521 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     8581 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     8641 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     8701 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     8761 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     8821 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     8881 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     8941 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     9001 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     9061 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     9121 ctcacgttaa gggattttgg t
//
