LOCUS       sequence-442658-        7333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expression of two genes in yeast with FLAG and MYC tags and ARG1 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7333)
  AUTHORS   Dong G, Nakai T, Matsuzaki T
  TITLE     A novel plasmid-based experimental system in Saccharomyces 
            cerevisiae that enables the introduction of 10 different plasmids 
            into cells.
  JOURNAL   FEBS Open Bio. 2024 Oct 10. doi: 10.1002/2211-5463.13893.
  PUBMED    39387713
REFERENCE   2  (bases 1 to 7333)
  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: "FEBS Open 
            Bio. 2024 Oct 10. doi: 10.1002/2211-5463.13893."
COMMENT     Sequence Label: pESC-ARG
FEATURES             Location/Qualifiers
     source          1..7333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2053..2508)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2190..2211)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2402..2421
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(2587..2752)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     2673..2697
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2900..2923)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        2970..3634
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    3186..3303
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3598..3621
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     3645..3664
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3645..3663
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3681..3710
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      3740..3929
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4001..4018)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4172..4760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4252..4271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4931..5722,5723..5791))
                     /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     5554..5573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5792..5896)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5923..7265
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(6886..6933)
                     /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)."
     primer_bind     7306..7324
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt caattcatga attgaattga aaagctgtag actaacatag
      241 cgctcttata taagagcgct atgttagtca ctactgacgg ctctccagtc atttatgtga
      301 ttttttagtg actcatgtcg catttggccc gtttttttcc gctgtcgcaa cctatttcca
      361 ttaacggtgc cgtatggaag agtcatttaa aggcaggaga gagagattac tcatcttcat
      421 tggatcagat tgatgactgc gtagcggcag atagtgtaat ctgagcagtt gcgagaccca
      481 gactggcact gtctcaatag tatattaatg ggcatacatt cgtactccct tgttcttgcc
      541 cacagttctc tctctcttta cttcttgtat cttgtctccc cattgtgcag cgataaggaa
      601 cattgttcta atatacacgg atacaaaaga aatacacata attgcataaa ataatgtcta
      661 agggaaaagt ttgtttggct tattctggtg gtttagatac ctccgtcatt ttggcttggc
      721 tactagacca aggctacgaa gttgtagctt tcatggctaa tgtagggcaa gaagaagatt
      781 tcgatgccgc caaggaaaag gccttgaaga tcggtgcctg caagttcgtt tgtgtggatt
      841 gtcgtgaaga ttttgtcaag gatattctat tcccagctgt acaggtcaac gctgtgtacg
      901 aagacgttta tctgttgggt acttctttgg caagacctgt tattgccaaa gcccaaattg
      961 acgtcgctaa acaggagggc tgtttcgcgg tctctcatgg ttgtaccggt aaaggtaatg
     1021 atcaaatcag attcgaattg tcattttacg ctctgaagcc agacgttaag tgtattacac
     1081 catggagaat gcctgaattt ttcgaaagat ttgctggcag aaaggatttg ttagactatg
     1141 ctgcacaaaa gggtattccc gtcgcccaaa ccaaggccaa gccatggtct actgacgaaa
     1201 accaagccca catttcttac gaggcaggta tcttggaaga cccagatacc accccaccaa
     1261 aggacatgtg gaaattgatc gtcgatccaa tggatgctcc ggaccaacca caagatttga
     1321 ccattgactt tgaacgtggt cttccagtca agttgaccta caccgacaac aagacttcca
     1381 aggaagtttc cgttaccaag cctttggatg ttttcttggc cgcatccaac ttagcaaggg
     1441 ccaacggtgt tggtagaatt gatattgtag aagatcgtta cattaacttg aaatccagag
     1501 gttgttacga acaggctcca ttgactgttt tgagaaaagc tcatgttgat ttggaaggtt
     1561 tgactttaga caaagaagtc cgtcaattga gagactcatt cgtcacacca aactactcca
     1621 gattgatata taacggttcc tacttcaccc cagagtgtga gtacatcaga tctatgatcc
     1681 aaccatccca aaatagcgtt aacggtactg tcagggttag actgtataag ggtaacgtca
     1741 tcattctggg cagatctaca aagactgaaa agttgtacga tccgacagaa tcctctatgg
     1801 atgagttgac cggtttctta cctaccgata ccaccggttt cattgccatc caggccatta
     1861 gaattaaaaa atacggtgaa tccaaaaaaa ccaaaggtga agagttgact ttgtaagtcc
     1921 gctagttcat cgcctcaaga tagataacga tctcttcctc cacctcctat ttctgcacac
     1981 tcttgtgatg taacccggga aatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     2041 taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2101 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2161 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2221 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2281 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     2341 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     2401 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     2461 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt
     2521 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2581 gaattggagc gacctcatgc tatacctgag aaagcaacct gacctacagg aaagagttac
     2641 tcaagaataa gaattttcgt tttaaaacct aagagtcact ttaaaatttg tatacactta
     2701 ttttttttat aacttattta ataataaaaa tcataaatca taagaaattc gcttatttag
     2761 aagtgtcaac aacgtatcta ccaacgattt gacccttttc catcttttcg taaatttctg
     2821 gcaaggtaga caagccgaca accttgattg gagacttgac caaacctctg gcgaagaatt
     2881 gttaattaag agctcagatc ttatcgtcgt catccttgta atccatcgat actagtgcgg
     2941 ccgcccttta gtgagggttg aattcgaatt ttcaaaaatt cttacttttt ttttggatgg
     3001 acgcaaagaa gtttaataat catattacat ggcattacca ccatatacat atccatatac
     3061 atatccatat ctaatcttac ttatatgttg tggaaatgta aagagcccca ttatcttagc
     3121 ctaaaaaaac cttctctttg gaactttcag taatacgctt aactgctcat tgctatattg
     3181 aagtacggat tagaagccgc cgagcgggtg acagccctcc gaaggaagac tctcctccgt
     3241 gcgtcctcgt cttcaccggt cgcgttcctg aaacgcagat gtgcctcgcg ccgcactgct
     3301 ccgaacaata aagattctac aatactagct tttatggtta tgaagaggaa aaattggcag
     3361 taacctggcc ccacaaacct tcaaatgaac gaatcaaatt aacaaccata ggatgataat
     3421 gcgattagtt ttttagcctt atttctgggg taattaatca gcgaagcgat gatttttgat
     3481 ctattaacag atatataaat gcaaaaactg cataaccact ttaactaata ctttcaacat
     3541 tttcggtttg tattacttct tattcaaatg taataaaagt atcaacaaaa aattgttaat
     3601 atacctctat actttaacgt caaggagaaa aaaccccgga tccgtaatac gactcactat
     3661 agggcccggg cgtcgacatg gaacagaagt tgatttccga agaagacctc gagtaagctt
     3721 ggtaccgcgg ctagctaaga tccgctctaa ccgaaaagga aggagttaga caacctgaag
     3781 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     3841 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     3901 tgcttgagaa ggttttggga cgctcgaaga tccagctgca ttaatgaatc ggccaacgcg
     3961 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4021 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4081 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4141 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4201 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4261 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4321 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4381 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4441 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4501 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4561 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4621 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4681 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4741 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4801 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4861 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4921 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4981 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5041 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5101 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5161 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5221 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5281 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5341 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5401 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5461 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5521 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5581 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5641 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5701 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5761 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5821 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5881 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaacgaagc atctgtgctt
     5941 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     6001 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt
     6061 gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     6121 tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa
     6181 tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag
     6241 catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt
     6301 ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca
     6361 taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat
     6421 tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt
     6481 gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc
     6541 ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga
     6601 ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa
     6661 acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt
     6721 aggttatata gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt
     6781 gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt
     6841 ttgaaagtgc gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt
     6901 tctagagaat aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc
     6961 gaaaatgcaa cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg
     7021 tgttgcctgt atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg
     7081 cgtacttata tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt
     7141 atcccattcc atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata
     7201 tgctgccact cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt
     7261 ggatcatact aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     7321 aggccctttc gtc
//
