LOCUS       sequence-45169-e        5478 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 5478)
  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 5478)
  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
FEATURES             Location/Qualifiers
     source          1..5478
                     /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      2675..2922
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2691..2709)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2999..3016)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3170..3758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3250..3269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3929..4720,4721..4789))
                     /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(4885..5478,1..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 agctgaaact
     2641 cgagcctgca ggcggccgca tctagagggc cgcatcatgt aattagttat gtcacgctta
     2701 cattcacgcc ctccccccac atccgctcta accgaaaagg aaggagttag acaacctgaa
     2761 gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt
     2821 caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc
     2881 ttgcttgaga aggttttggg acgctcgaag gctttaattt gcaagctgcg gccctgcatt
     2941 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3001 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3061 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3121 aaggccagca aaagcccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3181 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3241 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3301 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3361 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3421 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3481 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3541 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3601 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3661 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3721 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3781 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3841 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3901 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3961 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4021 cgatacggga gcgcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4081 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4141 gtcctgcaac tttatccgcc tccattcagt ctattaattg ttgccgggaa gctagagtaa
     4201 gtagttcgcc agttaatagt ttgcgcaacg ttgttggcat tgctacaggc atcgtggtgt
     4261 cactctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4321 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4381 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4441 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4501 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataatagtg
     4561 tatcacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4621 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4681 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4741 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4801 ttcaatgggt aataactgat ataattaaat tgaagctcta atttgtgagt ttagtataca
     4861 tgcatttact tataatacag ttttttagtt ttgctggccg catcttctca aatatgcttc
     4921 ccagcctgct tttctgtaac gttcaccctc taccttagca tcccttccct ttgcaaatag
     4981 tcctcttcca acaataataa tgtcagatcc tgtagagacc acatcatcca cggttctata
     5041 ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca taatcaacca
     5101 atcgtaacct tcatctcttc cacccatgtc tctttgagca ataaagccga taacaaaatc
     5161 tttgtcgctc ttcgcaatgt caacagtacc cttagtatat tctccagtag atagggagcc
     5221 cttgcatgac aattctgcta acatcaaaag gcctctaggt tcctttgtta cttcttctgc
     5281 cgcctgcttc aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc
     5341 tgcccattct gctattctgt atacacccgc agagtactgc aatttgactg tattaccaat
     5401 gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac ttggcggata atgcctttag
     5461 cggcttaact gtgccctc
//
