LOCUS       sequence-323986-        8344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express Dcp2 FL-H1 at endogenous level in S.cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8344)
  AUTHORS   Xing W, Muhlrad D, Parker R, Rosen MK
  TITLE     A quantitative inventory of yeast P body proteins reveals principles
            of composition and specificity.
  JOURNAL   Elife. 2020 Jun 19;9. pii: 56525. doi: 10.7554/eLife.56525.
  PUBMED    32553117
REFERENCE   2  (bases 1 to 8344)
  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"; journalName: "Elife."; 
            date: "2020-06-19"; volume: "9. pii: 56525. doi"; pages: " 
            10.7554/eLife.56525"
COMMENT     Sequence Label: Dcp2C Delta-5H-H1
FEATURES             Location/Qualifiers
     source          1..8344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    288..309
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(324..329,330..347,348..354)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    362..378
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     367..389
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     386..402
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     386..402
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     421..441
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        423..441
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2997..3707
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(3024..3052)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3647..3667
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3731..3918
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3786..3810)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3986..4002)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3987..4003)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4027..4046)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4028..4046)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4056..4073)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4056..4072)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4065..4087)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4213..4668
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4300..4319)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4510..4531
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4799..5602)
                     /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(5603..5818)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5842..5861)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5961..5983
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6021..6039)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6080..6583
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6620..6724
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6725..6793,6794..7585)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6943..6962)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7756..8344
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8245..8264
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      241 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      301 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      361 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
      421 gcaattaacc ctcactaaag ggaacaaaag ctggagctct cgtcgtaagg ctgacactgc
      481 agaagaaaag tcataatttg aatactagcc ggtatgaaac tgtgattgat taacctgggg
      541 ttacctaaag agaacataag taatactcat gacagaatca aaacacaata caaaatttat
      601 ccgaacctcg gcccgactgc cggctcgccg ggaaagggga caaccgcttc tatccgtcga
      661 ctaacttcat cggcccaatg gaagctatga tatggggatt tccattgagc cgatagcaat
      721 gtagggtaat actgttgcgt atatagtgat agttattgaa ttttattacc ctgcgggaat
      781 attgagacat cactaagcac gaattttacg tctgaggaaa gttgaatgat ggcaaaataa
      841 ccaggaaaaa caaatattga atccttgtga aggattccac agttgtttaa tcctccttaa
      901 gctcacttag tatcaattgt ctaaataata ttgctttgaa tctgaaaaaa ataaaagtac
      961 cttcgcatta gacaatgcct tcaatatttc catctctttc tgaaccgttt gctaacaata
     1021 agaatgttat accacctact atgccaatgg ctaacgtatt catgtcaaat cctcaattgt
     1081 ttgcgacaat gaatggccag ccttttgcac ctttcccatt tatgttacca ttaactaaca
     1141 atagtaatag cgctaaccct attccaactc cggtcccccc taattttaat gctcctccga
     1201 atccgatggc ttttggtgtt ccaaacatgc ataacctttc tggaccagca gtatctcaac
     1261 cgttttcctt gcctcctgct cctttaccga gggactctgg ttacagcagc tcctccgctg
     1321 cagcggcggc agctgcagca gctgcggcaa agcctgacag caacgtgcaa tcaagcaaaa
     1381 agccaaagct taaaatctta cagagaggaa cggacttgaa ttcaatcaag caaaacaata
     1441 atgatgaaac tgctcattca aacgctgcag ctgcggcagc tgcggcggca gcagcaacat
     1501 catcgcagaa gatacacgct tccaaaccag atacttcctt tttaccaaat gactccgtat
     1561 ctggtataca agatgcagaa tatgaagatt tcgagagtag ttcagatgaa gaggtggaga
     1621 cagctagaga tgaaagaaat tcattgaatg tagatattgg ggtgaacgtt atgccaagcg
     1681 aaaaagacag ccgaagaagt caaaaggaaa aaccaaggaa cgacgcaagc aaaacaaact
     1741 tgaacgcttc tgcagaatct aatagtgtag aatgggggcc tggtaaatct tcaccctcga
     1801 ctcagagtaa acagaattca tcagtaggaa tgcagaataa atatcgtcaa gaaattcaca
     1861 ttggtgatag tgatgcatat gaagtcttcg aaagcagttc tgacgaagaa gatggaaaaa
     1921 aattggaaga actggaacaa actcaagaca actccaaatt aattagccaa gatattttaa
     1981 aagagaacaa tttccaagac ggtgaagtgc ctcatagaga tatgcccact gaaagtaata
     2041 aatctataaa cgaaactgtc gggttatcat caacaacaaa tactgtaaaa aaggtacccg
     2101 cagctgcagc agcagcagca gctgaaacct ttgccagtct ggcgaatgat aaaaaagctt
     2161 ttgattcaag ctccaatgtg tcttcatcaa aagatttatt acagatgcta agaaacccaa
     2221 tttcctctac tgtatcttca aatcagcaat caccaaaatc tcaacatcta agtggggatg
     2281 aagaaatcat gatgatgttg aagaggaatt ctgtgtcaaa acctcaaaat agcgaagaaa
     2341 atgcatcaac ttcaagtata aacgatgcga atgcaagcga acttttggga atgttaaaac
     2401 aaaaggaaaa agacattaca gcaccaaaac agccctataa tgttgactca tactcgcaga
     2461 aaaattcagc taaaggagcg gcggctgctg cgaaaaagaa cgactctacc gggtaccctc
     2521 gcactgaggg tggaccgtca agtgaaatgt caacgtcaat gaaacgcaac gacgctacaa
     2581 ataatcaaga attagataag aattcaacag agttattaaa ctacttaaag cccaagcctc
     2641 ttaatgatgg gtatgagaat atttcaaata aggacagttc tcatgaatta ctgaatatat
     2701 tgcatggtaa taaaaacagc agcgccttca ataataatgt atatgcaact gacggatatt
     2761 ccttggcatc tgataacaac gaaaattcct caaataaagc agcagctgcg gcgcaaaaca
     2821 ggtctagtgc aataaatgag cccaacttcg acgtccgttc gaatggaact tcagggtcta
     2881 atgaattatt aagcattttg cataggaagg aagatcaatt gaaatcctat gcggaagaac
     2941 aattgaaatt gttgttgggt atcactaagg aggagcggat ccccgggtta attaacagta
     3001 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
     3061 atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta
     3121 cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca cttgtcacta
     3181 ctttcactta tggtgttcaa tgcttttcaa gatacccaga tcatatgaaa cggcatgact
     3241 ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg
     3301 acgggaacta caagacacgt gctgaagtca agtttgaagg tgataccctt gttaatagaa
     3361 tcgagttaaa aggtattgat tttaaagaag atggaaacat tcttggacac aaattggaat
     3421 acaactataa ctcacacaat gtatacatca tggcagacaa acaaaagaat ggaatcaaag
     3481 ttaacttcaa aattagacac aacattgaag atggaagcgt tcaactagca gaccattatc
     3541 aacaaaatac tccaattggc gatggtcctg tccttttacc agacaaccat tacctgtcca
     3601 cacaatctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt
     3661 ttgtaacagc tgctgggatt acacatggca tggatgaact atacaaatag ggcgcgccac
     3721 ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa
     3781 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt
     3841 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat
     3901 tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga tcccccgggc
     3961 tgcaggaatt cgatatcaag cttatcgata ccgtcgacct cgaggggggg cccggtaccc
     4021 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     4081 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     4141 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     4201 aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4261 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4321 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4381 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4441 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4501 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4561 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4621 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     4681 ttttctcctt acgcatctgt gcggtatttc acaccgcata gggtaataac tgatataatt
     4741 aaattgaagc tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt
     4801 agttttgctg gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac
     4861 cctctacctt agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag
     4921 atcctgtaga gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt
     4981 catctaaacc cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca
     5041 tgtctctttg agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag
     5101 tacccttagt atattctcca gtagataggg agcccttgca tgacaattct gctaacatca
     5161 aaaggcctct aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac
     5221 ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac
     5281 ccgcagagta ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga
     5341 gtaaaaaatt gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa
     5401 aatcagtcaa gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa
     5461 ctaactccag taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct
     5521 tttcgtgcat gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt
     5581 ccttatatgt agctttcgac atgatttatc ttcgtttcct gcaggttttt gttctgtgca
     5641 gttgggttaa gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata
     5701 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa
     5761 aaaaatttca aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa
     5821 ggtggtatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5881 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5941 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     6001 gaaacgcgcg agacgaaggg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     6061 aataatggtt tcttagacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa
     6121 tgatggaata atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat
     6181 tttagaaagt aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa
     6241 ggtttaaaaa atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc
     6301 agattaaata gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg
     6361 tgtgtggtct tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa
     6421 gagcaagata aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa
     6481 aacaaaaact attttttctt taatttcttt ttttactttc tatttttaat ttatatattt
     6541 atattaaaaa atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg
     6601 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6661 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6721 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6781 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6841 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6901 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6961 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     7021 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     7081 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     7141 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7201 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7261 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7321 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7381 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7441 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7501 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7561 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7621 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7681 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7741 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7801 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7861 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     7921 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7981 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     8041 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     8101 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     8161 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     8221 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     8281 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     8341 gaaa
//
