LOCUS       sequence-10222-e        5813 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 5813)
  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 5813)
  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-weakSC_GFP
FEATURES             Location/Qualifiers
     source          1..5813
                     /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      3010..3257
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3026..3044)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3334..3351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3505..4093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3585..3604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4264..5055,5056..5124))
                     /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(5220..5813,1..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 gaggaattct ttttttctag
     3001 agggccgcat catgtaatta gttatgtcac gcttacattc acgccctccc cccacatccg
     3061 ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta
     3121 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     3181 gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct
     3241 cgaaggcttt aatttgcaag ctgcggccct gcattaatga atcggccaac gcgcggggag
     3301 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3361 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3421 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagc ccaggaaccg
     3481 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3541 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3601 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3661 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3721 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3781 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3841 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3901 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     3961 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4021 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4081 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4141 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4201 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4261 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4321 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagcgct taccatctgg
     4381 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4441 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4501 tcagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4561 caacgttgtt ggcattgcta caggcatcgt ggtgtcactc tcgtcgtttg gtatggcttc
     4621 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4681 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4741 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4801 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4861 ttgctcttgc ccggcgtcaa tacgggataa tagtgtatca catagcagaa ctttaaaagt
     4921 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4981 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5041 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5101 gacacggaaa tgttgaatac tcatactctt cctttttcaa tgggtaataa ctgatataat
     5161 taaattgaag ctctaatttg tgagtttagt atacatgcat ttacttataa tacagttttt
     5221 tagttttgct ggccgcatct tctcaaatat gcttcccagc ctgcttttct gtaacgttca
     5281 ccctctacct tagcatccct tccctttgca aatagtcctc ttccaacaat aataatgtca
     5341 gatcctgtag agaccacatc atccacggtt ctatactgtt gacccaatgc gtctcccttg
     5401 tcatctaaac ccacaccggg tgtcataatc aaccaatcgt aaccttcatc tcttccaccc
     5461 atgtctcttt gagcaataaa gccgataaca aaatctttgt cgctcttcgc aatgtcaaca
     5521 gtacccttag tatattctcc agtagatagg gagcccttgc atgacaattc tgctaacatc
     5581 aaaaggcctc taggttcctt tgttacttct tctgccgcct gcttcaaacc gctaacaata
     5641 cctgggccca ccacaccgtg tgcattcgta atgtctgccc attctgctat tctgtataca
     5701 cccgcagagt actgcaattt gactgtatta ccaatgtcag caaattttct gtcttcgaag
     5761 agtaaaaaat tgtacttggc ggataatgcc tttagcggct taactgtgcc ctc
//
