LOCUS       sequence-317848-        7909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYC2/CT 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 7909)
  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 7909)
  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: pCFEX3-A2aGH
FEATURES             Location/Qualifiers
     source          1..7909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(72..863,864..932))
                     /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     695..714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     RBS             2052..2060
                     /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)"
     misc_feature    3252..3756
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     3786..3805
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3914..4355
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    3914..4031
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4326..4349
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     4387..4406
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4387..4405
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(5798..5800,5801..5803,5804..6514)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5798..5800,5801..5803,5804..6514)
                     /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(5843..5864)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6104..6123)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6451..6472
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6527..6556
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             6625..6666
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6676..6693
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      6727..6974
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6743..6761)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(7051..7068)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7222..7810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7302..7321)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
       61 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
      121 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
      181 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
      241 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
      301 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
      361 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
      421 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
      481 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
      541 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
      601 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
      661 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
      721 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
      781 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
      841 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
      901 ataagggcga cacggaaatg ttgaatactc atactctact cctcgtcaat tctcttagga
      961 ttcgattcac attcatcttt ttttagctac taataggata aattatagga atttataaca
     1021 aattgaaata tggcaggcag caaaattaaa attgtcttag ttttttagtg tatagaagtg
     1081 aatagctata taaagtatgt gtaaagttgg taacggaacg aaaaatagaa aaggatatta
     1141 catgggaaaa catgttgttt acggagaaat gaaaagtata ttgtattttg tacgagctaa
     1201 aagtacagtg ggaacaaagt cgattttgtt acatctacac tgttgttatc agatctcgag
     1261 ctattacaag tcttcttcag aaataagctt ttgttatcag gagacggtga ctgaggttcc
     1321 ttgaccccag tagtccatac catagtaaga acccgtacag taatagatac ccatgtcttc
     1381 aactcttaag ctaacaaatc gggttggtca atcctctagt ccaagcatag gatccgagag
     1441 ttatcaccaa taagcaatat gaaattgcaa tcgaaacagt ataataaata agcatgttaa
     1501 taaatgatag cttataccct tcattgataa acgtgctaat ggtactcaaa gtgccgcaaa
     1561 atccggatat taacgcagaa acaatatgac atgaattcaa cgagttcaca attggaacgt
     1621 ccgtagaaaa gtgtttttta ccacgttgca ccatggtaaa tatgccaatc aataaagtgg
     1681 cgaaaacgtt tgccaagaaa gtacccaatg gaaacttttt gtttgtttta ttaaacattt
     1741 ctgccaacca atatcttagg aaaccagcga aaattccaaa tagacatggt agtgtccact
     1801 tgcccctcga atagttctca taatacgcac acagcaccac aaacaaaata attaatggaa
     1861 aagctagtgc atacgcgagt ttatccacaa tatcaaaaaa tttcttaaaa aatggtgctg
     1921 gagttttgaa tttaaactct agaatatttt tttccatttc ttgtgtatca acatcaccag
     1981 cattttcctt tacagtgtct gagggagggg tatatggctt cgaaaacgaa ctagatccat
     2041 aagcaacaat aacctcctta ccaagttgcc taccgaaaat aagactaccc atggaaacca
     2101 taagatgaac gagcaagaca gataaaaact ccattatacc gtaagctctg ttcggcagtt
     2161 ttgtgtggtt agcgatatta ccgttagtta agtttgtaga gtgttcgaac atctctagca
     2221 gcatactaga aaaggaagac agggcaccac agtacccagt agttacacct aaaaacaaga
     2281 cttggtgatc tttcatccaa gtgtaagcat ttaaagattg cataatcccc attagcatgc
     2341 aggatgaaca attagaccag agaacggtcg gagcagtaac gaaggccgga gtgtaggtgg
     2401 aaagcgccgt tatagcctga cgtgtttcag tacctaatat gcagaacgtg gtgaatgtac
     2461 aaaaaacatg aatataatgg ctcaacttgt gatttgatat gaccggattg aaaatcattt
     2521 tgaattctcg ctggatatgc ctagaaatgc agtatcgatt tattttgaga aaatctgtca
     2581 actcgttact tctttgatag ggaaatcttt agttaaattt gtttaaaatt aagaaccagg
     2641 tctttagact atgccaaaag gtattggtgc ttaaaggttg ccgtaaactc taactctata
     2701 tatctctaag ccaatgttta agttgatgcg atgtgcatga tcacttcaca tttttttcat
     2761 gattttgttt tcttccttag ctactttttt ccgtcttggt ctgtgtaaaa aaaaaatcgc
     2821 ttcactgaag aaaaatataa ataaaaaaag gacatcagca agttcgtagt caataaatgt
     2881 gatgagtaat ttgcgataat ctaccccaac tcaaagcatt caattcaaat gaaagaagag
     2941 ttcgtgaatt ttaatctggc ttacagattg gatcatagct tctaatatat cttccttttt
     3001 attttaaagt gattaaaata tgtggatgtc tgcattaagg tgaaggcaga ctatagtgaa
     3061 aaatagtagc aaaggcttca tataatgaac tacctagttg tattcaattg gaatctgtga
     3121 acgcggcatt aaaagaatat ttgagctttt tttgattgta ttaacctgaa gctatcacgg
     3181 tttagactca ttaaaagaaa aatatccggt tgtatcggat aacaacacag gtcagctgat
     3241 gggtcctttt catcacgtgc tataaaaata attataattt aaatttttta atataaatat
     3301 ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa aaatagtttt tgttttccga
     3361 agatgtaaaa gactctaggg ggatcgccaa caaatactac cttttatctt gctcttcctg
     3421 ctctcaggta ttaatgccga attgtttcat cttgtctgtg tagaagacca cacacgaaaa
     3481 tcctgtgatt ttacatttta cttatcgtta atcgaatgta tatctattta atctgctttt
     3541 cttgtctaat aaatatatat gtaaagtacg ctttttgttg aaatttttta aacctttgtt
     3601 tatttttttt ttcttcattc cgtaactctt ctaccttctt tatttacttt ctaaaatcca
     3661 aatacaaaac ataaaaataa ataaacacag agtaaattcc caaattattc catcattaaa
     3721 agatacgagg cgcgtgtaag ttacaggcaa gcgatccgtc cgccggcgaa cgtggcgaga
     3781 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3841 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtggggatcg
     3901 atccactagt ccacggatta gaagccgccg agcgggtgac agccctccga aggaagactc
     3961 tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt gcctcgcgcc
     4021 gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg aagaggaaaa
     4081 attggcagta acctggcccc acaaaccttc aaatgaacga atcaaattaa caaccatagg
     4141 atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc gaagcgatga
     4201 tttttgatct attaacagat atataaatgc aaaaactgca taaccacttt aactaatact
     4261 ttcaacattt tcggtttgta ttacttctta ttcaaatgta ataaaagtat caacaaaaaa
     4321 ttgttaatat acctctatac tttaacgtca aggagaaaaa accccggatc ggactactag
     4381 cagctgtaat acgactcact atagggaata ttaagcttgg atcatgaagg ttttgattgt
     4441 cttgttggct atcttcgctg ctttgccctt ggctctagct caaccggtta tttctactac
     4501 cgtcggttcc gctgcagaag gctctttgga caagagagaa gctcggccgg acgtcatgcc
     4561 catcatgggc tcctcggtgt acatcacggt ggagctggcc attgctgtgc tggccatcct
     4621 gggcaatgtg ctggtgtgct gggccgtgtg gctcaacagc aacctgcaga acgtcaccaa
     4681 ctactttgtg gtgtcactgg cggcggccga catcgcagtg ggtgtgctcg ccatcccctt
     4741 tgccatcacc atcagcaccg ggttctgcgc tgcctgccac ggctgcctct tcattgcctg
     4801 cttcgtcctg gtcctcacgc agagctccat cttcagtctc ctggccatcg ccattgaccg
     4861 ctacattgcc atccgcatcc cgctccggta caatggcttg gtgaccggca cgagggctaa
     4921 gggcatcatt gccatctgct gggtgctgtc gtttgccatc ggcctgactc ccatgctagg
     4981 ttggaacaac tgcggtcagc caaaggaggg caagaaccac tcccagggct gcggggaggg
     5041 ccaagtggcc tgtctctttg aggatgtggt ccccatgaac tacatggtgt acttcaactt
     5101 ctttgcctgt gtgctggtgc ccctgctgct catgctgggt gtctatttgc ggatcttcct
     5161 ggcggcgcga cgacagctga agcagatgga gagccagcct ctgccggggg agcgggcacg
     5221 gtccacactg cagaaggagg tccatgctgc caagtcactg gccatcattg tggggctctt
     5281 tgccctctgc tggctgcccc tacacatcat caactgcttc actttcttct gccccgactg
     5341 cagccacgcc cctctctggc tcatgtacct ggccatcgtc ctctcccaca ccaattcggt
     5401 tgtgaatccc ttcatctacg cctaccgtat ccgcgagttc cgccagacct tccgcaagat
     5461 cattcgcagc cacgtcctga ggcagcaaga acctttcaag gcagctggca ccagtgcccg
     5521 ggtcttggca gctcatggca gtgacggaga gcaggtcagc ctccgtctca acggccaccc
     5581 gccaggagtg tgggccaacg gcagtgctcc ccaccctgag cggaggccca atggctacgc
     5641 cctggggctg gtgagtggag ggagtgccca agagtcccag gggaacacgg gcctcccaga
     5701 cgtggagctc cttagccatg agctcaaggg agtgtgccca gagccccctg gcctagatga
     5761 ccccctggcc caggatggag caggagtgtc cccgcggatg gtgagcaagg gcgaggagct
     5821 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     5881 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     5941 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     6001 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     6061 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     6121 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     6181 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     6241 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     6301 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     6361 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     6421 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     6481 cgggatcact ctcggcatgg acgagctgta caagcacgtg tcatcacatc atcatcatca
     6541 tcatcatcat catcattaat aacttaagag ttgaagacat gggtatctat tactgtacgg
     6601 ctcgagtcta gagggccctt cgaaggtaag cctatcccta accctctcct cggtctcgat
     6661 tctacgcgta ccggtcatca tcaccatcac cattgagttt aaacccgctg atcctagagg
     6721 gccgcatcat gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc
     6781 taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag
     6841 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac
     6901 gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga
     6961 aggctttaat ttgcaagctg cggccctgca ttaatgaatc ggccaacgcg cggggagagg
     7021 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     7081 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7141 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7201 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7261 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7321 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7381 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7441 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7501 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7561 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7621 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     7681 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7741 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7801 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7861 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacc
//
