LOCUS       sequence-454040-        7354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expression of two genes in yeast with V5 and HA tags and ARG1 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7354)
  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 7354)
  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 (V5 and HA)
FEATURES             Location/Qualifiers
     source          1..7354
                     /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(2897..2938)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     promoter        2991..3655
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    3207..3324
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3619..3642
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     3666..3685
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3666..3684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3702..3731
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      3761..3950
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4022..4039)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4193..4781)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4273..4292)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4952..5743,5744..5812))
                     /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     5575..5594
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5813..5917)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5944..7286
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(6907..6954)
                     /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     7327..7345
                     /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 agctcaggta gaatccagac ccagcagagg gttagggata ggcttaccag
     2941 atctcatcga tactagtgcg gccgcccttt agtgagggtt gaattcgaat tttcaaaaat
     3001 tcttactttt tttttggatg gacgcaaaga agtttaataa tcatattaca tggcattacc
     3061 accatataca tatccatata catatccata tctaatctta cttatatgtt gtggaaatgt
     3121 aaagagcccc attatcttag cctaaaaaaa ccttctcttt ggaactttca gtaatacgct
     3181 taactgctca ttgctatatt gaagtacgga ttagaagccg ccgagcgggt gacagccctc
     3241 cgaaggaaga ctctcctccg tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga
     3301 tgtgcctcgc gccgcactgc tccgaacaat aaagattcta caatactagc ttttatggtt
     3361 atgaagagga aaaattggca gtaacctggc cccacaaacc ttcaaatgaa cgaatcaaat
     3421 taacaaccat aggatgataa tgcgattagt tttttagcct tatttctggg gtaattaatc
     3481 agcgaagcga tgatttttga tctattaaca gatatataaa tgcaaaaact gcataaccac
     3541 tttaactaat actttcaaca ttttcggttt gtattacttc ttattcaaat gtaataaaag
     3601 tatcaacaaa aaattgttaa tatacctcta tactttaacg tcaaggagaa aaaaccccgg
     3661 atccgtaata cgactcacta tagggcccgg gcgtcgacat ggaacagaag ttgatttccg
     3721 aagaagacct cgagtaagct tggtaccgcg gctagctaag atccgctcta accgaaaagg
     3781 aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat
     3841 taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat
     3901 gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag atccagctgc
     3961 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4021 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4081 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4141 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4201 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4261 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4321 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4381 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4441 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4501 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4561 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4621 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4681 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4741 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4801 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4861 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4921 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4981 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5041 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5101 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5161 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5221 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5281 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5341 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5401 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5461 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5521 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5581 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5641 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5701 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5761 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5821 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5881 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5941 ctgaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt
     6001 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta
     6061 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc
     6121 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag
     6181 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag
     6241 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata
     6301 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt
     6361 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta
     6421 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc
     6481 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt
     6541 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt
     6601 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct
     6661 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca
     6721 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag
     6781 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca
     6841 gtccggtgcg tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag
     6901 cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggaa cttcaaagcg
     6961 tttccgaaaa cgagcgcttc cgaaaatgca acgcgagctg cgcacataca gctcactgtt
     7021 cacgtcgcac ctatatctgc gtgttgcctg tatatatata tacatgagaa gaacggcata
     7081 gtgcgtgttt atgcttaaat gcgtacttat atgcgtctat ttatgtagga tgaaaggtag
     7141 tctagtacct cctgtgatat tatcccattc catgcggggt atcgtatgct tccttcagca
     7201 ctacccttta gctgttctat atgctgccac tcctcaattg gattagtctc atccttcaat
     7261 gctatcattt cctttgatat tggatcatac taagaaacca ttattatcat gacattaacc
     7321 tataaaaata ggcgtatcac gaggcccttt cgtc
//
