LOCUS       sequence-323989-        8542 bp    DNA     circular SYN 15-APR-2025
DEFINITION  express wild type Dcp2 Delta-N(243-970) at endogenous level in 
            S.cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8542)
  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 8542)
  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 Delta-N
FEATURES             Location/Qualifiers
     source          1..8542
                     /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             3195..3905
                     /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(3222..3250)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3845..3865
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3929..4116
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3984..4008)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4184..4200)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4185..4201)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4225..4244)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4226..4244)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4254..4271)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4254..4270)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4263..4285)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4411..4866
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4498..4517)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4708..4729
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4997..5800)
                     /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(5801..6016)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6040..6059)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6159..6181
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6219..6237)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6278..6781
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6818..6922
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6923..6991,6992..7783)
                     /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(7141..7160)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7954..8542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8443..8462
                     /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 gacatcgaca tcgcgaatga atgaagatca attgaaatcc tatgcggaag
     1021 aacaattgaa attgttgttg ggtatcacta aggaggagca gattgatccc ggtagagagt
     1081 tgctgaatat gttacatact gcagtgcaag ctaacagtaa taataatgcg gtctccaacg
     1141 gacaggtacc ctcgagccaa gagcttcagc atttgaaaga gcaatcagga gaacacaacc
     1201 aacagaagga tcagcagtca tcgttttctt ctcaacaaca accttcaata tttccatctc
     1261 tttctgaacc gtttgctaac aataagaatg ttataccacc tactatgcca atggctaacg
     1321 tattcatgtc aaatcctcaa ttgtttgcga caatgaatgg ccagcctttt gcacctttcc
     1381 catttatgtt accattaact aacaatagta atagcgctaa ccctattcca actccggtcc
     1441 cccctaattt taatgctcct ccgaatccga tggcttttgg tgttccaaac atgcataacc
     1501 tttctggacc agcagtatct caaccgtttt ccttgcctcc tgctccttta ccgagggact
     1561 ctggttacag cagctcctcc cctgggcagt tgttagatat actaaattcg aaaaagcctg
     1621 acagcaacgt gcaatcaagc aaaaagccaa agcttaaaat cttacagaga ggaacggact
     1681 tgaattcaat caagcaaaac aataatgatg aaactgctca ttcaaactct caagctttgc
     1741 tagatttgtt gaaaaaacca acatcatcgc agaagataca cgcttccaaa ccagatactt
     1801 cctttttacc aaatgactcc gtatctggta tacaagatgc agaatatgaa gatttcgaga
     1861 gtagttcaga tgaagaggtg gagacagcta gagatgaaag aaattcattg aatgtagata
     1921 ttggggtgaa cgttatgcca agcgaaaaag acagccgaag aagtcaaaag gaaaaaccaa
     1981 ggaacgacgc aagcaaaaca aacttgaacg cttctgcaga atctaatagt gtagaatggg
     2041 ggcctggtaa atcttcaccc tcgactcaga gtaaacagaa ttcatcagta ggaatgcaga
     2101 ataaatatcg tcaagaaatt cacattggtg atagtgatgc atatgaagtc ttcgaaagca
     2161 gttctgacga agaagatgga aaaaaattgg aagaactgga acaaactcaa gacaactcca
     2221 aattaattag ccaagatatt ttaaaagaga acaatttcca agacggtgaa gtgcctcata
     2281 gagatatgcc cactgaaagt aataaatcta taaacgaaac tgtcgggtta tcatcaacaa
     2341 caaatactgt aaaaaaggta cccaaagtta aaatactaaa acgaggtgaa acctttgcca
     2401 gtctggcgaa tgataaaaaa gcttttgatt caagctccaa tgtgtcttca tcaaaagatt
     2461 tattacagat gctaagaaac ccaatttcct ctactgtatc ttcaaatcag caatcaccaa
     2521 aatctcaaca tctaagtggg gatgaagaaa tcatgatgat gttgaagagg aattctgtgt
     2581 caaaacctca aaatagcgaa gaaaatgcat caacttcaag tataaacgat gcgaatgcaa
     2641 gcgaactttt gggaatgtta aaacaaaagg aaaaagacat tacagcacca aaacagccct
     2701 ataatgttga ctcatactcg cagaaaaatt cagctaaagg attgttgaat attttgaaaa
     2761 agaacgactc taccgggtac cctcgcactg agggtggacc gtcaagtgaa atgtcaacgt
     2821 caatgaaacg caacgacgct acaaataatc aagaattaga taagaattca acagagttat
     2881 taaactactt aaagcccaag cctcttaatg atgggtatga gaatatttca aataaggaca
     2941 gttctcatga attactgaat atattgcatg gtaataaaaa cagcagcgcc ttcaataata
     3001 atgtatatgc aactgacgga tattccttgg catctgataa caacgaaaat tcctcaaata
     3061 aactactaaa tatgctgcaa aacaggtcta gtgcaataaa tgagcccaac ttcgacgtcc
     3121 gttcgaatgg aacttcaggg tctaatgaat tattaagcat tttgcatagg aagcggatcc
     3181 ccgggttaat taacagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg
     3241 aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg
     3301 caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat
     3361 ggccaacact tgtcactact ttcacttatg gtgttcaatg cttttcaaga tacccagatc
     3421 atatgaaacg gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa
     3481 ctatattttt caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg
     3541 atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc
     3601 ttggacacaa attggaatac aactataact cacacaatgt atacatcatg gcagacaaac
     3661 aaaagaatgg aatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggaagcgttc
     3721 aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
     3781 acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagacc
     3841 acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat
     3901 acaaataggg cgcgccactt ctaaataagc gaatttctta tgatttatga tttttattat
     3961 taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa
     4021 acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata
     4081 gtatgaggtc gctcttattg accacacctc taccggcaga tccgctaggg ataacagggt
     4141 aatatagatc ccccgggctg caggaattcg atatcaagct tatcgatacc gtcgacctcg
     4201 agggggggcc cggtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     4261 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     4321 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     4381 caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg
     4441 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4501 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4561 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4621 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4681 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4741 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4801 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     4861 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatagg
     4921 gtaataactg atataattaa attgaagctc taatttgtga gtttagtata catgcattta
     4981 cttataatac agttttttag ttttgctggc cgcatcttct caaatatgct tcccagcctg
     5041 cttttctgta acgttcaccc tctaccttag catcccttcc ctttgcaaat agtcctcttc
     5101 caacaataat aatgtcagat cctgtagaga ccacatcatc cacggttcta tactgttgac
     5161 ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt cataatcaac caatcgtaac
     5221 cttcatctct tccacccatg tctctttgag caataaagcc gataacaaaa tctttgtcgc
     5281 tcttcgcaat gtcaacagta cccttagtat attctccagt agatagggag cccttgcatg
     5341 acaattctgc taacatcaaa aggcctctag gttcctttgt tacttcttct gccgcctgct
     5401 tcaaaccgct aacaatacct gggcccacca caccgtgtgc attcgtaatg tctgcccatt
     5461 ctgctattct gtatacaccc gcagagtact gcaatttgac tgtattacca atgtcagcaa
     5521 attttctgtc ttcgaagagt aaaaaattgt acttggcgga taatgccttt agcggcttaa
     5581 ctgtgccctc catggaaaaa tcagtcaaga tatccacatg tgtttttagt aaacaaattt
     5641 tgggacctaa tgcttcaact aactccagta attccttggt ggtacgaaca tccaatgaag
     5701 cacacaagtt tgtttgcttt tcgtgcatga tattaaatag cttggcagca acaggactag
     5761 gatgagtagc agcacgttcc ttatatgtag ctttcgacat gatttatctt cgtttcctgc
     5821 aggtttttgt tctgtgcagt tgggttaaga atactgggca atttcatgtt tcttcaacac
     5881 tacatatgcg tatatatacc aatctaagtc tgtgctcctt ccttcgttct tccttctgtt
     5941 cggagattac cgaatcaaaa aaatttcaag gaaaccgaaa tcaaaaaaaa gaataaaaaa
     6001 aaaatgatga attgaaaagg tggtatggtg cactctcagt acaatctgct ctgatgccgc
     6061 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     6121 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     6181 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     6241 ataggttaat gtcatgataa taatggtttc ttagacggat cgcttgcctg taacttacac
     6301 gcgcctcgta tcttttaatg atggaataat ttgggaattt actctgtgtt tatttatttt
     6361 tatgttttgt atttggattt tagaaagtaa ataaagaagg tagaagagtt acggaatgaa
     6421 gaaaaaaaaa taaacaaagg tttaaaaaat ttcaacaaaa agcgtacttt acatatatat
     6481 ttattagaca agaaaagcag attaaataga tatacattcg attaacgata agtaaaatgt
     6541 aaaatcacag gattttcgtg tgtggtcttc tacacagaca agatgaaaca attcggcatt
     6601 aatacctgag agcaggaaga gcaagataaa aggtagtatt tgttggcgat ccccctagag
     6661 tcttttacat cttcggaaaa caaaaactat tttttcttta atttcttttt ttactttcta
     6721 tttttaattt atatatttat attaaaaaat ttaaattata attattttta tagcacgtga
     6781 tgaaaaggac ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6841 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6901 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6961 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7021 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7081 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7141 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7201 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7261 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7321 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7381 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7441 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7501 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7561 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7621 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     7681 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     7741 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7801 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7861 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7921 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7981 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     8041 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     8101 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     8161 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     8221 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     8281 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     8341 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     8401 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     8461 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     8521 aggggggcgg agcctatgga aa
//
