LOCUS       sequence-10219-e       10099 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 10099)
  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 10099)
  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-Ago1
FEATURES             Location/Qualifiers
     source          1..10099
                     /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             4756..4797
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4807..4824
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4845..5092
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4861..4879)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5169..5186)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5340..5928)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5420..5439)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6099..6890,6891..6959))
                     /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(7055..7858)
                     /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(7859..8079)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      8678..9558
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(9627..10082)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     9976..9995
                     /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 ttatgtcatc caattcggag gagaacagtc
      541 aagttcctcc ccttgatgcc accgctgctg caactaaacc aaaaaaggcc aagaaaccaa
      601 aggttaagaa gccaaaggat tccgctgagg catcttcatc accagctgct gagggcactg
      661 ctgaagccaa gccaaaaaag gctaagaaat caaagactaa gaagtcaaag gagtctgccg
      721 aggtatcgcc agctccagct gatgaaacta cttctgctgg tgtagatgct aaacctaaga
      781 aggctaaaaa gtcaaaggtc aagaagccaa aggattctac tgaatcatct cctgctccaa
      841 gtaatgaacc tccagccgcc gaagttgccg ccgaagatgc aaagtctaaa aaggttaaga
      901 aaccaaaggc caagaaacca aaggaatcta ctgaatcatc tcctgctcca ggtcaggaag
      961 ccgctgctac tgaaggtgcc acagaagata aacctaagaa ggttaagaaa tcaaaggcta
     1021 agaaggcaaa ggaatctgtt gaatcatctc ctgccgccac tgaatctgtc tctgagaaga
     1081 ctgcaaagaa atctaagaag ccaaaggcta agaaatctac ttctcctgaa acaactgaag
     1141 aaataactga agaatctacc gaatcaaagg aaaagaagac aaagacaaag aaaccaaagg
     1201 aaaagaagtc atctccatct actgccactt cgactgctgc ttcaaaacca gttacttcca
     1261 ttgcgggagt caccattcca ggcaagactt ttgatttgac agatgttcct ccatctaaac
     1321 ctgctccaaa gatggtcccg gaagcctata aattacaaac tagagtggat tacggtacta
     1381 agggtaccaa agtggacgtc ttgactaatc atatactact atctgttggt gatgatgtcc
     1441 ctcaagatga aagagcctct caattggatc catggtggaa atctgcattt gtatatacct
     1501 ataacattac ttttgccgtt ccacagagta attcaccacg taaaggtcca gcgcctgctt
     1561 tatccaaacc aaagaaatac gaattggtag aatctttgtt tcacgaggat gaaaccttgt
     1621 tcaaatataa agatcgtatc tccttcaatg gtgaagatac tctatactct catgtcccat
     1681 tggaggaatt cactttattt gatggttgtt gggatgtcag taacaagcaa aagaaaagaa
     1741 gacaagaagt tgttggtctc aacagtagag ccaaggaaat taatgacttg gctgctcaag
     1801 ttactttaaa atttgctgat aaagtcccat tgggtgatat ttataaggct accacttcaa
     1861 aggatccaga agaacaagaa aataagatgg ctaacgctga taaggttgct ttattgtctt
     1921 tgatgggtgt caaattctta aacacaaagg aacaaatctt ccaacttaac ggtaataaat
     1981 tctttatttt taatgaacac gcaattgcta ctccattcca aattggtggg ttcttaatgc
     2041 atgggttcac agtttcccta cgatatgctt acgggtccgt tttgttaaat actgttaacg
     2101 tctgcctccc atttgttaag tggactaaat atttaccagg tgatgctaaa ttcaaggaaa
     2161 atgaaaagac tcaatacagt ttgttagatt ggattattga atgtatgcac caggccagtg
     2221 ctcaaagagg tcaaaagcta agaggccctc catctgccaa ggatatcaat tttttcattg
     2281 acaagaatag agacattaaa gatctattga agggtttgaa atgttacaga ccatacatta
     2341 attattctgt taacccagat ggaactccaa aaccaccaaa aaagatgcaa gctaagggta
     2401 tcgttggttt cgtaagagag actccagatt ctatgaaatt cagaacttta ccaagtaaca
     2461 tggaaaagaa tggtgtacca aaaccaggtg aaaaggaaat catggtcaca acaaccgcct
     2521 actttgccaa gaagtatgac atcaaattga aatacccaga tgttaaaatg gtgagtctag
     2581 gtggttcgaa tgttgtccca gctgaatgtt taaccattgt tccaggtcag aaattaaagg
     2641 gtttggttta tgacgaaaaa gccgttattg atttcactgc tttaagacca agtgaaaaat
     2701 tcagagccat caccaacttg gcattacctg ccattaagag agctttatca actgaagaag
     2761 aaaatgctaa ggctccacat gattctggtt acactttcat gaaggtccca tctcgtgtca
     2821 ttgacgctcc tgtggttcaa ttcaagaaca ctacagtcac atatgtggat aaaccatttg
     2881 gtactaagaa tggtaagaat aatcatgaag aaactaaggg taattggaat ttaaaggatc
     2941 ataaattcat tactgttcca aaggaaccaa tgcacttgag agctatcttt attaatgatt
     3001 ctgataagtc tccaccagtt tctgtcatgg atgaattgaa ggcctctttg agcaaattcg
     3061 ctgaagatgt cgctgatgtc ggtgttaact ttgacgtatc catggcccca attttaatta
     3121 acaattttaa tgccccaata aagaaggtta ctggtggctt tggtggtaga ggtggacgtg
     3181 gtggtagagg cggccgtggt ggccgtggtg gtagaggagg tcgtggtggt ttcggcggtg
     3241 gccgtggtga aactacttat gaattaactc caggtgaaga aaaactacgt cacttgttag
     3301 ccaacgtccc agaaaagact tatgtcttat tcgtcttggg tcgtggggat gactctgcta
     3361 tttacaacag attgaaatat ttagctgatt tgacgtacgg tgtcattaac aattgtgtta
     3421 tttggaacaa gttcagaaag tgttccactc aatacaatgt taatgtggta atgaagatga
     3481 acttgaagtt agaaggtgct aaccactcct tatgtgcaga agatataaac ttactgaagg
     3541 atgaaaaatc aggattacca ttcatgattt taggtgctga tgttacacat tatccagaaa
     3601 aggatcagaa ttctatctct gcattggtgg gttcttttga tgacaaattt gctcaattcc
     3661 caggttcata tatgttacaa agtggtccag gtgaagaaat aattgctggt atcggtaaca
     3721 tggttctaca gagattgaaa ctgtatcaaa agcacaacaa tggcaaacta cctccaaaga
     3781 ttctattcta cagagatggt gtttctgaat cccaattctc acaaattgtt caaattgaag
     3841 ttaagggttt gaaacaagca ttgaagaaat tcggtagtga attaaacaag ggtgtcaact
     3901 ataacccatc tgtcaccaca atttgtgttg tcaagagaaa tcaaattaga tttatgccac
     3961 ttgagcaaaa tgctataaat gaaaagggtg aagttgctgc cgttcaatca ttcgagaacg
     4021 ttatgccagg tactgttgtt gatcgtggta tcacctctag tgctcacttc gatttcttct
     4081 tgcaatctca acaaccattg aagggtactg gtgttccatg tcattactgg tgtatctacg
     4141 acgaaaatca attcaattct gattacttgc aacaagttac tcacgctttg tgttacttat
     4201 tcggtagatc gagtacaagt attaaggtcg catctcctgt ttattacgct gatttattgt
     4261 gtgaacgtgg tgctgcattc ttcaaggcta actttgagct tgctcaatac gaattttcta
     4321 aggagaggaa gaacagagat gacgttatac cgaccggtaa attactacaa cctgttcata
     4381 agaatgtcac tgacatcatg tactacatat gaacaaagag tttatgtgct gcctactttt
     4441 gcttattttt gtctattccc cgtactattt ctatcaactt tctagcttaa atgtatgttt
     4501 aaaaacatct attattaata tcctattctt ttatttaatt ttattattgc aaaatttggt
     4561 gtcagcttcg cgaacaatcg ttcgatgagg aataaaaaaa aatttaaaat gaaaaaaaaa
     4621 aaattattcc ttaagctata aacttttagt agactcagta caagggaaat caccaaatat
     4681 tataatatat aatatccacg atgtctgata tgttgccgtt agccaaaggg cgagcttcga
     4741 ggtcacccat tcgaaggtaa gcctatccct aaccctctcc tcggtctcga ttctacgcgt
     4801 accggtcatc atcaccatca ccattgagtt tctagagggc cgcatcatgt aattagttat
     4861 gtcacgctta cattcacgcc ctccccccac atccgctcta accgaaaagg aaggagttag
     4921 acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt
     4981 atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat
     5041 actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt gcaagctgcg
     5101 gccctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5161 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5221 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5281 atgtgagcaa aaggccagca aaagcccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5341 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5401 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5461 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5521 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5581 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5641 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5701 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5761 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5821 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5881 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5941 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6001 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6061 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6121 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6181 tagataacta cgatacggga gcgcttacca tctggcccca gtgctgcaat gataccgcga
     6241 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6301 cgcagaagtg gtcctgcaac tttatccgcc tccattcagt ctattaattg ttgccgggaa
     6361 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttggcat tgctacaggc
     6421 atcgtggtgt cactctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6481 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6541 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6601 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6661 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6721 gataatagtg tatcacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6781 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6841 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6901 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6961 ctcttccttt ttcaatgggt aataactgat ataattaaat tgaagctcta atttgtgagt
     7021 ttagtataca tgcatttact tataatacag ttttttagtt ttgctggccg catcttctca
     7081 aatatgcttc ccagcctgct tttctgtaac gttcaccctc taccttagca tcccttccct
     7141 ttgcaaatag tcctcttcca acaataataa tgtcagatcc tgtagagacc acatcatcca
     7201 cggttctata ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca
     7261 taatcaacca atcgtaacct tcatctcttc cacccatgtc tctttgagca ataaagccga
     7321 taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc cttagtatat tctccagtag
     7381 atagggagcc cttgcatgac aattctgcta acatcaaaag gcctctaggt tcctttgtta
     7441 cttcttctgc cgcctgcttc aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat
     7501 tcgtaatgtc tgcccattct gctattctgt atacacccgc agagtactgc aatttgactg
     7561 tattaccaat gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac ttggcggata
     7621 atgcctttag cggcttaact gtgccctcca tggaaaaatc agtcaagata tccacatgtg
     7681 tttttagtaa acaaattttg ggacctaatg cttcaactaa ctccagtaat tccttggtgg
     7741 tacgaacatc caatgaagca cacaagtttg tttgcttttc gtgcatgata ttaaatagct
     7801 tggcagcaac aggactagga tgagtagcag cacgttcctt atatgtagct ttcgacatga
     7861 tttatcttcg tttcctgcag gtttttgttc tgtgcagttg ggttaagaat actgggcaat
     7921 ttcatgtttc ttcaacacta catatgcgta tatataccaa tctaagtctg tgctccttcc
     7981 ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa atttcaaaga aaccgaaatc
     8041 aaaaaaaaga ataaaaaaaa aatgatgaat tgaattgaaa agctagctta tcgatgataa
     8101 gctgtcaaag atgagaatta attccacgga ctatagacta tactagatac tccgtctact
     8161 gtacgataca cttccgctca ggtccttgtc ctttaacgag gccttaccac tcttttgtta
     8221 ctctattgat ccagctcagc aaaggcagtg tgatctaaga ttctatcttc gcgatgtagt
     8281 aaaactagct agaccgagaa agagactaga aatgcaaaag gcacttctac aatggctgcc
     8341 atcattatta tccgatgtga cgctgcagct tctcaatgat attcgaatac gctttgagga
     8401 gatacagcct aatatccgac aaactgtttt acagatttac gatcgtactt gttacccatc
     8461 attgaatttt gaacatccga acctgggagt tttccctgaa acagatagta tatttgaacc
     8521 tgtataataa tatatagtct agcgctttac ggaagacaat gtatgtattt cggttcctgg
     8581 agaaactatt gcatctattg cataggtaat cttgcacgtc gcatccccgg ttcattttct
     8641 gcgtttccat cttgcacttc aatagcatat ctttgttaac gaagcatctg tgcttcattt
     8701 tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     8761 ttttacagaa cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc
     8821 atttttgtaa aacaaaaatg caacgcgacg agagcgctaa tttttcaaac aaagaatctg
     8881 agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc
     8941 tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca
     9001 tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt
     9061 gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata
     9121 aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt
     9181 tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt
     9241 gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt
     9301 ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt
     9361 cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac
     9421 ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag
     9481 gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt
     9541 ggaagcggta ttcgcaatgg gaagctccac cccggttgat aatcagaaaa gccccaaaaa
     9601 caggaagatt gtataagcaa atatttaaat tgtaaacgtt aatattttgt taaaattcgc
     9661 gttaaatttt tgttaaatca gctcattttt taacgaatag cccgaaatcg gcaaaatccc
     9721 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ccaacaagag
     9781 tccactatta aagaacgtgg actccaacgt caaagggcga aaaagggtct atcagggcga
     9841 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     9901 agtaaatcgg aagggtaaac ggatgccccc atttagagct tgacggggaa agccggcgaa
     9961 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggg gctagggcgg tgggaagtgt
    10021 aggggtcacg ctgggcgtaa ccaccacacc cgccgcgctt aatggggcgc tacagggcgc
    10081 gtggggatga tccactagt
//
