LOCUS       sequence-10220-e        7720 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 7720)
  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 7720)
  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: pYES2.1-Dcr1
FEATURES             Location/Qualifiers
     source          1..7720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..443
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2..119
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     414..437
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     475..494
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        475..493
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2377..2418
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2428..2445
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2466..2713
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2482..2500)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2790..2807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2961..3549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3041..3060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3720..4511,4512..4580))
                     /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(4676..5479)
                     /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"
     promoter        complement(5480..5700)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      6299..7179
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(7248..7703)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     7597..7616
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt
       61 cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
      121 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
      181 ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga
      241 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
      301 taacagatat ataaatgcaa aaactgcata accactttaa ctaatacttt caacattttc
      361 ggtttgtatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt gttaatatac
      421 ctctatactt taacgtcaag gagaaaaaac cccggatcgg actactagca gctgtaatac
      481 gactcactat agggaatatt aagctcgccc ttcaccatga atagagaaaa aagcgccgat
      541 ctaagtaaaa catcagatgc tccatacaat gaactcgatg ccaaaaattt aagaaatttc
      601 tataaggttc aaaatgcatg tgcgcaattg agggaatcca taaaagtaat ttacgaaaac
      661 gggttatcat ccgatcaatt gaatactatg gcgaaacatg ggaacgacct agaaaaatct
      721 attgcaaaca gtcctgctat gtccgtagca agttgcttga accaagttag acccacccta
      781 gacatcaaga atatctttga tcattataaa tttgaaaata attcatcacc agttgatcct
      841 tatgtccatt atcccgtatg ttctgatcaa aatttggaaa atttggcttt cattcataga
      901 tctttgccga atatgaatgt caaattgact gaattgcaaa agaccgtaat gagtaatgaa
      961 cgtttggaat ttttaggtga tagttggctt ggcgccttag tggcgtatat tatttataag
     1021 aaatatcctt atgctaatga aggtgcctta tcaaagatga aggaagctat cgttaacaac
     1081 aacaatttgg aaaagatctg tgaaaaactt ggctttaaag aaagattaaa ggaaaatatt
     1141 ccacgttctt caatgaaaat caaagataga ttaaccaaga attacgcaga ctgtgtggaa
     1201 gcctacattg gcgcattggt tattgataga ttttcaacag aatttaatga tgttgctctt
     1261 tggcttgaag aactatctga ggaacatttc attgaactag gacctatgat ggtcaaagag
     1321 cctttaaaca aaaatgcaaa gggtgaacta ggtgcgttct tacaatttaa taatattgga
     1381 gccaaaatat catataaaag attgaatgat aaatctccgt ttaaggtaga agtaagattg
     1441 gggaataatt tgttaggtat tggagatggt tcaaatgtga gagaagctga acagagagcc
     1501 gctatggaag cacttgctca accgaaattg attcaaaaat attctctaca tgatattgaa
     1561 ctagaaagag gcatgattga agaagcagat aatgttcccc aattactcaa aagtgcggaa
     1621 attcccaatc aaacaccaca aagtcatcat tctatcacag aagaaattga tgaagaagaa
     1681 ggtcttacat tcccccattc gaatccattt ttgaaaggta caccaccagc agttttacct
     1741 gaaattgtaa ctaatcaaat tccacaggct tctgcggatc atcaagaaca aaatgtgcct
     1801 gacactgata aaattgtgaa tgatgttatg gaacgtatga gtaagatact gtcggtgatg
     1861 gtttctgagg ctgtttcaaa tgcacttggt agaccagctc caaaaaatgc acctatccct
     1921 atcccagtaa caaccccagt ctcaccgcta gcaacatccc cagtctcagt ctcagtccca
     1981 gcgccagctc cagccgtagc tgtcacccca tcagttgtga ataatccacc acaacccaaa
     2041 atggtcacac aaccggtaag cacaaaccta aataaaacgc ctgaagttgc acttttaaat
     2101 tctgtgtatg ataaagaagc ctcagggaga ctttatgcat tgttaggtaa atataaattg
     2161 tatccagaat ataataccga gcaactagga atgacagatt tttatacggt atgctttata
     2221 aagggtgttg gtgtagaaat tggtaaaggc catggtagaa gtaaaaagat atcccaacat
     2281 aaatctgcag aagatgcttt gaatggaaaa gcactgaagg aatacttgag gcattgcaac
     2341 aatctgaagg gcgagcttcg aggtcaccca ttcgaaggta agcctatccc taaccctctc
     2401 ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt ttctagaggg
     2461 ccgcatcatg taattagtta tgtcacgctt acattcacgc cctcccccca catccgctct
     2521 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt tttttatagt
     2581 tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacagacg
     2641 cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg gacgctcgaa
     2701 ggctttaatt tgcaagctgc ggccctgcat taatgaatcg gccaacgcgc ggggagaggc
     2761 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2821 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2881 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaagcccag gaaccgtaaa
     2941 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3001 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3061 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3121 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3181 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3241 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3301 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3361 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     3421 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3481 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3541 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3601 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3661 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3721 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3781 gttgcctgac tccccgtcgt gtagataact acgatacggg agcgcttacc atctggcccc
     3841 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3901 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccattcag
     3961 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4021 gttgttggca ttgctacagg catcgtggtg tcactctcgt cgtttggtat ggcttcattc
     4081 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4141 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4201 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4261 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4321 tcttgcccgg cgtcaatacg ggataatagt gtatcacata gcagaacttt aaaagtgctc
     4381 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4441 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4501 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4561 cggaaatgtt gaatactcat actcttcctt tttcaatggg taataactga tataattaaa
     4621 ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca gttttttagt
     4681 tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct
     4741 ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata atgtcagatc
     4801 ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct cccttgtcat
     4861 ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt ccacccatgt
     4921 ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac
     4981 ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct aacatcaaaa
     5041 ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta acaatacctg
     5101 ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg tatacacccg
     5161 cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta
     5221 aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc atggaaaaat
     5281 cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat gcttcaacta
     5341 actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt gtttgctttt
     5401 cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca gcacgttcct
     5461 tatatgtagc tttcgacatg atttatcttc gtttcctgca ggtttttgtt ctgtgcagtt
     5521 gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt atatatacca
     5581 atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc gaatcaaaaa
     5641 aatttcaaag aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa ttgaattgaa
     5701 aagctagctt atcgatgata agctgtcaaa gatgagaatt aattccacgg actatagact
     5761 atactagata ctccgtctac tgtacgatac acttccgctc aggtccttgt cctttaacga
     5821 ggccttacca ctcttttgtt actctattga tccagctcag caaaggcagt gtgatctaag
     5881 attctatctt cgcgatgtag taaaactagc tagaccgaga aagagactag aaatgcaaaa
     5941 ggcacttcta caatggctgc catcattatt atccgatgtg acgctgcagc ttctcaatga
     6001 tattcgaata cgctttgagg agatacagcc taatatccga caaactgttt tacagattta
     6061 cgatcgtact tgttacccat cattgaattt tgaacatccg aacctgggag ttttccctga
     6121 aacagatagt atatttgaac ctgtataata atatatagtc tagcgcttta cggaagacaa
     6181 tgtatgtatt tcggttcctg gagaaactat tgcatctatt gcataggtaa tcttgcacgt
     6241 cgcatccccg gttcattttc tgcgtttcca tcttgcactt caatagcata tctttgttaa
     6301 cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca
     6361 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta
     6421 ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgac gagagcgcta
     6481 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
     6541 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
     6601 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
     6661 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
     6721 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
     6781 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
     6841 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
     6901 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
     6961 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
     7021 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
     7081 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
     7141 atacttttga gcaatgtttg tggaagcggt attcgcaatg ggaagctcca ccccggttga
     7201 taatcagaaa agccccaaaa acaggaagat tgtataagca aatatttaaa ttgtaaacgt
     7261 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaacgaata
     7321 gcccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     7381 tgttccagtt tccaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     7441 aaaaagggtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     7501 ggggtcgagg tgccgtaaag cagtaaatcg gaagggtaaa cggatgcccc catttagagc
     7561 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     7621 ggctagggcg gtgggaagtg taggggtcac gctgggcgta accaccacac ccgccgcgct
     7681 taatggggcg ctacagggcg cgtggggatg atccactagt
//
