LOCUS       sequence-10223-e        6082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6082)
  AUTHORS   Drinnenberg IA, Weinberg DE, Xie KT, Mower JP, Wolfe KH, Fink GR, 
            Bartel DP
  TITLE     RNAi in budding yeast.
  JOURNAL   Science. 2009 Oct 23;326(5952):544-550. doi: 
            10.1126/science.1176945. Epub 2009 Sep 10.
  PUBMED    19745116
REFERENCE   2  (bases 1 to 6082)
  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"; doi: 
            "10.1126/science.1176945"; journalName: "Science"; date: 
            "2009-10-23- 23"; volume: "326"; issue: "5952"; pages: "544-550"
COMMENT     Sequence Label: plp-strongSC_GFP
FEATURES             Location/Qualifiers
     source          1..6082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(211..431)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     promoter        2163..2604
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2163..2280
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2575..2598
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     terminator      3279..3526
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3295..3313)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3603..3620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3774..4362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3854..3873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4533..5324,5325..5393))
                     /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"
     CDS             complement(join(5489..6082,1..210))
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
ORIGIN
        1 catggaaaaa tcagtcaaga tatccacatg tgtttttagt aaacaaattt tgggacctaa
       61 tgcttcaact aactccagta attccttggt ggtacgaaca tccaatgaag cacacaagtt
      121 tgtttgcttt tcgtgcatga tattaaatag cttggcagca acaggactag gatgagtagc
      181 agcacgttcc ttatatgtag ctttcgacat gatttatctt cgtttcctgc aggtttttgt
      241 tctgtgcagt tgggttaaga atactgggca atttcatgtt tcttcaacac tacatatgcg
      301 tatatatacc aatctaagtc tgtgctcctt ccttcgttct tccttctgtt cggagattac
      361 cgaatcaaaa aaatttcaaa gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga
      421 attgaattga aaagctagcg atcccttatc aaatatggta cttaaaagtg aagatttctc
      481 ttttttttgg aatttatttt actttttgat cttcttttac tattgtaaaa ggtaaaatta
      541 gataaataaa aattctatgc taatcaatat aaataatttt tcagtttcac taaaaacctc
      601 taagtacata caatggactt catgatcaac agctattcta ttgttcacat tgttgcccct
      661 gtatttaaca aaaaaaaggc cttctcagaa atgtatccct ttcaataatg atgcccttga
      721 tagcgccgct aaagaagaaa gagttatgag aaataattct ttgacatttc aagtacagta
      781 tagacagaaa acaaatttag gggtgggaaa gagtttacag aaaaagcata ttgaggaaaa
      841 catctggtaa ataaaagctg tgagttaccg gttttttccc tctccagcct gatactatga
      901 gaaaataccc atgatttttc tcatagtatc gaagaagtgc agacaaggtc gaaaacgcct
      961 tctatttcca cctgtcacaa aatccaatcc actatataat gttaattgat ctaatctagt
     1021 aaaaaatata gtcaaggaac caatactgtc aaatgggagt actaacgtac aatgaaaaac
     1081 aagataaagg tgaacagccc aattgaagaa aggaaagaaa acaagtctac cttttgctgt
     1141 tttaggataa atacttttta taaaagcggt cttaaaagtc cagaaagtta caccttcttc
     1201 cctaaagaac atgtattata accacgactc caagcaccta cctctatacg ttttaatgaa
     1261 tatacgtaag atgtgtttaa cgctacttga taggatactc caatctgaga gctctatact
     1321 tcactttcaa atacgtaatt actatgcaca agggaaacat atcggtttcc ctgcataaat
     1381 aagttcttac accttctctc cacgtaatcg atattacgca aatacgtact ttctcagtaa
     1441 taaccgccca cgccaagagg acccccacat tataacaagc aatctccggt ataatttgcg
     1501 gggccggtga aactccgaga aaaaaggaaa agaaaaaaaa agaggcgccg ttggcacaaa
     1561 acccagacac cggaattccg cattttctat tacaggaatc caccacccac gcatctatag
     1621 gtataatttt ggcagccccc aactaaaaaa tgatcagttt cctctacccc taaaggtttt
     1681 caccaacagg gttaataccc tcataatagt actcagtata agctccattc atcaggaaag
     1741 caagagacca cttaaatata aagtaaagaa gctttcaaga gaggctccat tactatggga
     1801 aggggactgt cgtggacgaa atacgccaca attttgccga gaaggtcatt agtatgtcca
     1861 agaaacccta ggtgtaaagt cgggaaatcc gaatctccga ttttggaggg gcccatgccc
     1921 tacttttttt cgccaggggt gaaattccaa acccgtgcgc gttcttggaa tttgacagcg
     1981 cattgagtat gtgctgcgta ttcccactat catgacgcgc cctttatctg ggaaaaatgg
     2041 aactggatgc tgaaatattt cactctcaga tcacatatcc caaatcctgt gagtgaattg
     2101 tttggtcagg cgaccaaaca ggaatatgga atagattcta ttctctggat tctacactag
     2161 tacggattag aagccgccga gcgggtgaca gccctccgaa ggaagactct cctccgtgcg
     2221 tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg cctcgcgccg cactgctccg
     2281 aacaataaag attctacaat actagctttt atggttatga agaggaaaaa ttggcagtaa
     2341 cctggcccca caaaccttca aatgaacgaa tcaaattaac aaccatagga tgataatgcg
     2401 attagttttt tagccttatt tctggggtaa ttaatcagcg aagcgatgat ttttgatcta
     2461 ttaacagata tataaatgca aaaactgcat aaccacttta actaatactt tcaacatttt
     2521 cggtttgtat tacttcttat tcaaatgtaa taaaagtatc aacaaaaaat tgttaatata
     2581 cctctatact ttaacgtcaa ggagaaaaaa ccccggatcg gactactagc agctgagaca
     2641 cgtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt
     2701 gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac
     2761 aatgtataca tcatggcaga caaacaaaag aatggaatca aagttaactt caaaattaga
     2821 cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt
     2881 ggcgatggcc ctgtcctttt accagacaac ctcgagaaaa tcaggtgtgt tggaactttt
     2941 ttcaaacctt actaaacatt gaaactaatt ggtaaagata gaggaattcg ttgtctggta
     3001 aaaggacagg gccatcgcca attggagtat tttgttgata atggtctgct agttgaacgc
     3061 ttccatcttc aatgttgtgt ctaattttga agttaacttt gattccattc ttttgtttgt
     3121 ctgccatgat gtatacattg tgtgagttat agttgtattc caatttgtgt ccaagaatgt
     3181 ttccatcttc tttaaaatca atacctttta actcgattct attaacaagg gtatcacctt
     3241 caaacttgac ttcagcacgt gtcttctaga gggccgcatc atgtaattag ttatgtcacg
     3301 cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     3361 tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat
     3421 atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa
     3481 aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcaagc tgcggccctg
     3541 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3601 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3661 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3721 gcaaaaggcc agcaaaagcc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3781 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3841 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3901 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3961 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4021 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4081 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4141 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4201 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4261 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4321 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4381 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4441 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4501 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4561 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4621 actacgatac gggagcgctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4681 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4741 agtggtcctg caactttatc cgcctccatt cagtctatta attgttgccg ggaagctaga
     4801 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg gcattgctac aggcatcgtg
     4861 gtgtcactct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4921 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4981 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5041 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5101 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5161 agtgtatcac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5221 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5281 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5341 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5401 ctttttcaat gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta
     5461 tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg
     5521 cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa
     5581 atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc
     5641 tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca
     5701 accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa
     5761 aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca gtagataggg
     5821 agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt
     5881 ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa
     5941 tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac
     6001 caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct
     6061 ttagcggctt aactgtgccc tc
//
