LOCUS       sequence-10221-e        7007 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 7007)
  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 7007)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..7007
                     /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             join(546..548,549..551,552..1262)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(591..612)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(852..871)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1199..1220
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          1348..1577
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1560..1579
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1664..1705
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1715..1732
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1753..2000
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1769..1787)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2077..2094)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2248..2836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2328..2347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3007..3798,3799..3867))
                     /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(3963..4766)
                     /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(4767..4987)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5586..6466
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6535..6990)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     6884..6903
                     /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 ttatggctag cggccgctct agcccgggcg
      541 gatccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      601 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      661 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      721 ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
      781 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      841 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      901 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
      961 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1021 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1081 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1141 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1201 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1261 agaagcttta agaattcagt ggatccacta gtaacggccg ccagtgtgct ggaattaatt
     1321 cgctgtctgc gagggccagc tgttggggtg agtactccct ctcaaaagcg ggcatgactt
     1381 ctgcgctaag attgtcagtt tccaaaaacg aggaggattt gatattcacc tggcccgcgg
     1441 tgatgccttt gagggtggcc gcgtccatct ggtcagaaaa gacaatcttt ttgttgtcaa
     1501 gcttgaggtg tggcaggctt gagatctggc catacacttg agtgacaatg acatccactt
     1561 tgcctttctc tccacaggtg tccactccca ggtccaactg caggtcgagc atgcatctag
     1621 ggcggccaat tccaagggcg agcttcgagg tcacccattc gaaggtaagc ctatccctaa
     1681 ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgagtttc
     1741 tagagggccg catcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat
     1801 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     1861 ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
     1921 acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac
     1981 gctcgaaggc tttaatttgc aagctgcggc cctgcattaa tgaatcggcc aacgcgcggg
     2041 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2101 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2161 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa agcccaggaa
     2221 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2281 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2341 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2401 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2461 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2521 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2581 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2641 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     2701 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2761 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2821 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2881 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2941 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3001 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3061 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagc gcttaccatc
     3121 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3181 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3241 cattcagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3301 gcgcaacgtt gttggcattg ctacaggcat cgtggtgtca ctctcgtcgt ttggtatggc
     3361 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3421 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3481 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3541 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3601 gagttgctct tgcccggcgt caatacggga taatagtgta tcacatagca gaactttaaa
     3661 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3721 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3781 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3841 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatgggtaa taactgatat
     3901 aattaaattg aagctctaat ttgtgagttt agtatacatg catttactta taatacagtt
     3961 ttttagtttt gctggccgca tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt
     4021 tcaccctcta ccttagcatc ccttcccttt gcaaatagtc ctcttccaac aataataatg
     4081 tcagatcctg tagagaccac atcatccacg gttctatact gttgacccaa tgcgtctccc
     4141 ttgtcatcta aacccacacc gggtgtcata atcaaccaat cgtaaccttc atctcttcca
     4201 cccatgtctc tttgagcaat aaagccgata acaaaatctt tgtcgctctt cgcaatgtca
     4261 acagtaccct tagtatattc tccagtagat agggagccct tgcatgacaa ttctgctaac
     4321 atcaaaaggc ctctaggttc ctttgttact tcttctgccg cctgcttcaa accgctaaca
     4381 atacctgggc ccaccacacc gtgtgcattc gtaatgtctg cccattctgc tattctgtat
     4441 acacccgcag agtactgcaa tttgactgta ttaccaatgt cagcaaattt tctgtcttcg
     4501 aagagtaaaa aattgtactt ggcggataat gcctttagcg gcttaactgt gccctccatg
     4561 gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac aaattttggg acctaatgct
     4621 tcaactaact ccagtaattc cttggtggta cgaacatcca atgaagcaca caagtttgtt
     4681 tgcttttcgt gcatgatatt aaatagcttg gcagcaacag gactaggatg agtagcagca
     4741 cgttccttat atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg
     4801 tgcagttggg ttaagaatac tgggcaattt catgtttctt caacactaca tatgcgtata
     4861 tataccaatc taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa
     4921 tcaaaaaaat ttcaaagaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg
     4981 aattgaaaag ctagcttatc gatgataagc tgtcaaagat gagaattaat tccacggact
     5041 atagactata ctagatactc cgtctactgt acgatacact tccgctcagg tccttgtcct
     5101 ttaacgaggc cttaccactc ttttgttact ctattgatcc agctcagcaa aggcagtgtg
     5161 atctaagatt ctatcttcgc gatgtagtaa aactagctag accgagaaag agactagaaa
     5221 tgcaaaaggc acttctacaa tggctgccat cattattatc cgatgtgacg ctgcagcttc
     5281 tcaatgatat tcgaatacgc tttgaggaga tacagcctaa tatccgacaa actgttttac
     5341 agatttacga tcgtacttgt tacccatcat tgaattttga acatccgaac ctgggagttt
     5401 tccctgaaac agatagtata tttgaacctg tataataata tatagtctag cgctttacgg
     5461 aagacaatgt atgtatttcg gttcctggag aaactattgc atctattgca taggtaatct
     5521 tgcacgtcgc atccccggtt cattttctgc gtttccatct tgcacttcaa tagcatatct
     5581 ttgttaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa
     5641 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc
     5701 tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgacgag
     5761 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     5821 gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc
     5881 ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct
     5941 ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc
     6001 tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact
     6061 gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct
     6121 ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca
     6181 ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa
     6241 tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt
     6301 tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa
     6361 gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag
     6421 caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatggga agctccaccc
     6481 cggttgataa tcagaaaagc cccaaaaaca ggaagattgt ataagcaaat atttaaattg
     6541 taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     6601 acgaatagcc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     6661 tgagtgttgt tccagtttcc aacaagagtc cactattaaa gaacgtggac tccaacgtca
     6721 aagggcgaaa aagggtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     6781 gttttttggg gtcgaggtgc cgtaaagcag taaatcggaa gggtaaacgg atgcccccat
     6841 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     6901 gagcgggggc tagggcggtg ggaagtgtag gggtcacgct gggcgtaacc accacacccg
     6961 ccgcgcttaa tggggcgcta cagggcgcgt ggggatgatc cactagt
//
