LOCUS       sequence-323987-        7243 bp    DNA     circular SYN 15-APR-2025
DEFINITION  express Dcp2 300 at endogenous level in S.cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7243)
  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 7243)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: Dcp2 300
FEATURES             Location/Qualifiers
     source          1..7243
                     /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             1896..2606
                     /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(1923..1951)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2546..2566
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2630..2817
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2685..2709)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2885..2901)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2886..2902)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2926..2945)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2927..2945)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2955..2972)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2955..2971)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2964..2986)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3112..3567
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3199..3218)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3409..3430
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3698..4501)
                     /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(4502..4717)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4741..4760)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4860..4882
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4920..4938)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4979..5482
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5519..5623
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5624..5692,5693..6484)
                     /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(5842..5861)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6655..7243
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7144..7163
                     /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 gacaatgtca ctgccgctac gacacgcatt ggagaacgtt acttctgttg
     1021 atagaatttt agaggactta ttagtacgtt ttattataaa ttgtccgaat gaagatttat
     1081 cgagtgtcga gagagagtta tttcattttg aagaagcctc atggttttac acggatttca
     1141 tcaaattgat gaatccaact ttaccctccc taaagattaa atcatttgct caattgatca
     1201 taaaactatg tcctctggtt tggaaatggg acataagagt ggatgaggca ctccagcaat
     1261 tctccaagta taagaaaagt ataccggtga ggggcgctgc catatttaac gagaacctga
     1321 gtaaaatttt attggtacag ggtactgaat cggattcttg gtcattccca agggggaaga
     1381 tatctaaaga tgaaaatgac atagattgtt gcattagaga agtgaaagaa gaaattggtt
     1441 tcgatttgac ggactatatt gacgacaacc aattcattga aagaaatatt caaggtaaaa
     1501 attacaaaat atttttgata tctggtgttt cagaagtctt caattttaaa cctcaagtta
     1561 gaaatgaaat tgataagata gaatggttcg attttaagaa aatttctaaa acaatgtaca
     1621 aatcaaatat caagtattat ctgattaatt ccatgatgag acccttatca atgtggttaa
     1681 ggcatcagag gcaaataaaa aatgaagatc aattgaaatc ctatgcggaa gaacaattga
     1741 aattgttgtt gggtatcact aaggaggagc agattgatcc cggtagagag ttgctgaata
     1801 tgttacatac tgcagtgcaa gctaacagta ataataatgc ggtctccaac ggacaggtac
     1861 cctcgagcca agagcggatc cccgggttaa ttaacagtaa aggagaagaa cttttcactg
     1921 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     1981 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     2041 ctggaaaact acctgttcca tggccaacac ttgtcactac tttcacttat ggtgttcaat
     2101 gcttttcaag atacccagat catatgaaac ggcatgactt tttcaagagt gccatgcccg
     2161 aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg
     2221 ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt
     2281 ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac tcacacaatg
     2341 tatacatcat ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa attagacaca
     2401 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     2461 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     2521 atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta
     2581 cacatggcat ggatgaacta tacaaatagg gcgcgccact tctaaataag cgaatttctt
     2641 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     2701 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     2761 caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct ctaccggcag
     2821 atccgctagg gataacaggg taatatagat cccccgggct gcaggaattc gatatcaagc
     2881 ttatcgatac cgtcgacctc gagggggggc ccggtaccca attcgcccta tagtgagtcg
     2941 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     3001 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     3061 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     3121 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3181 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3241 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3301 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3361 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3421 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3481 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3541 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     3601 cggtatttca caccgcatag ggtaataact gatataatta aattgaagct ctaatttgtg
     3661 agtttagtat acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc
     3721 tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc
     3781 cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat
     3841 ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg
     3901 tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc
     3961 cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag
     4021 tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg
     4081 ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg
     4141 cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga
     4201 ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg
     4261 ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat
     4321 gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg
     4381 tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata
     4441 gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca
     4501 tgatttatct tcgtttcctg caggtttttg ttctgtgcag ttgggttaag aatactgggc
     4561 aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct
     4621 tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa
     4681 atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag gtggtatggt gcactctcag
     4741 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4801 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4861 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4921 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgga
     4981 tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa tttgggaatt
     5041 tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta aataaagaag
     5101 gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa
     5161 aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag atatacattc
     5221 gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt ctacacagac
     5281 aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa aaggtagtat
     5341 ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta ttttttcttt
     5401 aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa tttaaattat
     5461 aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg gaaatgtgcg
     5521 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5581 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5641 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5701 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5761 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5821 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5881 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5941 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6001 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6061 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6121 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6181 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6241 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6301 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6361 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6421 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6481 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6541 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6601 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6661 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6721 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6781 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6841 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6901 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6961 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7021 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7081 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7141 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7201 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaa
//
