LOCUS       sequence-323895-        8278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express Dcp2C Delta-5H  at endogenous level in S.cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8278)
  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 8278)
  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
FEATURES             Location/Qualifiers
     source          1..8278
                     /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             2931..3641
                     /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(2958..2986)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3581..3601
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3665..3852
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3720..3744)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3920..3936)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3921..3937)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3961..3980)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3962..3980)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3990..4007)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3990..4006)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3999..4021)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4147..4602
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4234..4253)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4444..4465
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4733..5536)
                     /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(5537..5752)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5776..5795)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5895..5917
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5955..5973)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6014..6517
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6554..6658
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6659..6727,6728..7519)
                     /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(6877..6896)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7690..8278
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8179..8198
                     /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 cataggaagc ggatccccgg gttaattaac agtaaaggag
     2941 aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc
     3001 acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta
     3061 aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc actactttca
     3121 cttatggtgt tcaatgcttt tcaagatacc cagatcatat gaaacggcat gactttttca
     3181 agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga
     3241 actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt
     3301 taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact
     3361 ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc aaagttaact
     3421 tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa
     3481 atactccaat tggcgatggt cctgtccttt taccagacaa ccattacctg tccacacaat
     3541 ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa
     3601 cagctgctgg gattacacat ggcatggatg aactatacaa atagggcgcg ccacttctaa
     3661 ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa aaaaaataag
     3721 tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat tcttgagtaa
     3781 ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc ttattgacca
     3841 cacctctacc ggcagatccg ctagggataa cagggtaata tagatccccc gggctgcagg
     3901 aattcgatat caagcttatc gataccgtcg acctcgaggg ggggcccggt acccaattcg
     3961 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     4021 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4081 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4141 gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4201 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4261 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4321 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4381 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4441 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4501 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4561 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     4621 ccttacgcat ctgtgcggta tttcacaccg catagggtaa taactgatat aattaaattg
     4681 aagctctaat ttgtgagttt agtatacatg catttactta taatacagtt ttttagtttt
     4741 gctggccgca tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt tcaccctcta
     4801 ccttagcatc ccttcccttt gcaaatagtc ctcttccaac aataataatg tcagatcctg
     4861 tagagaccac atcatccacg gttctatact gttgacccaa tgcgtctccc ttgtcatcta
     4921 aacccacacc gggtgtcata atcaaccaat cgtaaccttc atctcttcca cccatgtctc
     4981 tttgagcaat aaagccgata acaaaatctt tgtcgctctt cgcaatgtca acagtaccct
     5041 tagtatattc tccagtagat agggagccct tgcatgacaa ttctgctaac atcaaaaggc
     5101 ctctaggttc ctttgttact tcttctgccg cctgcttcaa accgctaaca atacctgggc
     5161 ccaccacacc gtgtgcattc gtaatgtctg cccattctgc tattctgtat acacccgcag
     5221 agtactgcaa tttgactgta ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa
     5281 aattgtactt ggcggataat gcctttagcg gcttaactgt gccctccatg gaaaaatcag
     5341 tcaagatatc cacatgtgtt tttagtaaac aaattttggg acctaatgct tcaactaact
     5401 ccagtaattc cttggtggta cgaacatcca atgaagcaca caagtttgtt tgcttttcgt
     5461 gcatgatatt aaatagcttg gcagcaacag gactaggatg agtagcagca cgttccttat
     5521 atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg tgcagttggg
     5581 ttaagaatac tgggcaattt catgtttctt caacactaca tatgcgtata tataccaatc
     5641 taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat
     5701 ttcaaggaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg aaaaggtggt
     5761 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     5821 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5881 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5941 cgcgagacga aggggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     6001 ggtttcttag acggatcgct tgcctgtaac ttacacgcgc ctcgtatctt ttaatgatgg
     6061 aataatttgg gaatttactc tgtgtttatt tatttttatg ttttgtattt ggattttaga
     6121 aagtaaataa agaaggtaga agagttacgg aatgaagaaa aaaaaataaa caaaggttta
     6181 aaaaatttca acaaaaagcg tactttacat atatatttat tagacaagaa aagcagatta
     6241 aatagatata cattcgatta acgataagta aaatgtaaaa tcacaggatt ttcgtgtgtg
     6301 gtcttctaca cagacaagat gaaacaattc ggcattaata cctgagagca ggaagagcaa
     6361 gataaaaggt agtatttgtt ggcgatcccc ctagagtctt ttacatcttc ggaaaacaaa
     6421 aactattttt tctttaattt ctttttttac tttctatttt taatttatat atttatatta
     6481 aaaaatttaa attataatta tttttatagc acgtgatgaa aaggacccag gtggcacttt
     6541 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6601 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6661 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6721 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6781 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6841 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6901 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6961 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     7021 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     7081 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     7141 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     7201 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     7261 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     7321 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     7381 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     7441 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     7501 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     7561 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7621 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7681 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7741 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7801 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7861 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7921 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7981 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     8041 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     8101 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     8161 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     8221 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaa
//
