LOCUS       sequence-107360-        7254 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast two-hybrid bait vector expressing LexA-PPTR-1 fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7254)
  AUTHORS   Wang JT, Smith J, Chen BC, Schmidt H, Rasoloson D, Paix A, Lambrus 
            BG, Calidas D, Betzig E, Seydoux G
  TITLE     Regulation of RNA granule dynamics by phosphorylation of 
            serine-rich, intrinsically disordered proteins in C. elegans.
  JOURNAL   Elife. 2014 Dec 23;4. doi: 10.7554/eLife.04591.
  PUBMED    25535836
REFERENCE   2  (bases 1 to 7254)
  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: "2014-12-23"; volume: "4. doi"; pages: " 10.7554/eLife.04591"
COMMENT     Sequence Label: pJW3.42
FEATURES             Location/Qualifiers
     source          1..7254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..406
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             join(433..684,685..1038)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     primer_bind     976..997
                     /label=LexA
                     /note="LexA DNA binding domain, forward primer"
     terminator      2727..2914
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2782..2806)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2967..3641)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3642..3743)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3757..3773)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3757..3773)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3770..3792)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3781..3797
                     /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)."
     promoter        complement(join(3805..3811,3812..3829,3830..3835))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3850..3871
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3988..4005)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4159..4747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4239..4258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4916..5731)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4916..5731)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5639..5658
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5723..5742
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     5876..5894
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5932..5954)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6054..6073
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      6090..7254
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gcttgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc
       61 atatccgcaa tgacaaaaaa aatgatggaa gacactaaag gaaaaaatta acgacaaaga
      121 cagcaccaac agatgtcgtt gttccagagc tgatgagggg tatcttcgaa cacacgaaac
      181 tttttccttc cttcattcac gcacactact ctctaatgag caacggtata cggccttcct
      241 tccagttact tgaatttgaa ataaaaaaag tttgctgtct tgctatcaag tataaataga
      301 cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt
      361 ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa gcttgaatta
      421 attccgggcg gaatgaaagc gttaacggcc aggcaacaag aggtgtttga tctcatccgt
      481 gatcacatca gccagacagg tatgccgccg acgcgtgcgg aaatcgcgca gcgtttgggg
      541 ttccgttccc caaacgcggc tgaagaacat ctgaaggcgc tggcacgcaa aggcgttatt
      601 gaaattgttt ccggcgcatc acgcgggatt cgtctgttgc aggaagagga agaagggttg
      661 ccgctggtag gtcgtgtggc tgccggtgaa ccacttctgg cgcaacagca tattgaaggt
      721 cattatcagg tcgatccttc cttattcaag ccgaatgctg atttcctgct gcgcgtcagc
      781 gggatgtcga tgaaagatat cggcattatg gatggtgact tgctggcagt gcataaaact
      841 caggatgtac gtaacggtca ggtcgttgtc gcacgtattg atgacgaggt taccgttaag
      901 cgcctgaaaa aacagggcaa taaagtcgaa ctgttgccag aaaatagcga gtttaaacca
      961 attgtcgtag atcttcgtca gcagagcttc accattgaag ggctggcggt tggggttatt
     1021 cgcaacggcg actggctgga attcatgcac ggaagcgggc acagtctaac gggcgctccg
     1081 catcaaattc cgccgccacg gacacaggga gccgccaccg gtggccaaca actctcggcg
     1141 acagctaatc aatttgttga taaaatcgat ccgttccata ataaaagagg aacttctaga
     1201 cgattgcgca tcaacaacag cagccgatat aatgttgatt cggctcaaga actcgtacaa
     1261 ttggcgctca tcaaagatac tgctgcaaat gagcaaccgg ctctcgttat cgaaaagctc
     1321 gtccaatgtc agcacgtatt cgatttctat gatccagtag ctcaactgaa atgcaaagaa
     1381 atcaaaagag cagctctaaa tgagctcatc gatcacatta catcgacaaa aggagcaatt
     1441 gttgagacaa tttatccggc tgtcattaaa atggttgcca agaatatatt ccgtgtactt
     1501 ccaccatcag aaaattgtga atttgatcca gaagaggatg aaccgacttt agaagtctcg
     1561 tggccacatt tacagcttgt ctacgagctc ttcctgagat tccttgagtc gccagatttc
     1621 caagcttcaa tcggtaaaaa gtatattgat cagagatttg tgctcaagct gctcgattta
     1681 ttcgattcgg aggatccacg tgaacgagac tttttgaaga cggttcttca tcgtatttac
     1741 ggaaaatttt tgggccttcg ggcttttatt cgcaaacata tcaataatat gtttttgaga
     1801 ttcgtatacg aaacggactc attcaacggt gtcggtgagc ttctcgagat tcttggctca
     1861 attataaacg gatttgctct tccattgaag caagagcaca aggttttcct tgtaaaagtt
     1921 cttctaccat tgcacaaacc gaaatgctta tcactgtatc atgcacagct cgcctactgc
     1981 gtcgttcaat tcatcgaaaa agactcatca ctgactccac aagtttttga ggctctgctc
     2041 aaattttggc ctcgaacatg tagcagcaag gaggttatgt tcctgggtga agtggaggaa
     2101 attctcgaca ttatcgaacc ggaacaattc aaaaagatta tcgatccatt attccgtcaa
     2161 ttggccaaat gtgtcagtag tccacatttt caagttgctg aacgggctct ctacttttgg
     2221 aataatgaat atatattgtc attaattgag gatacaagca gtttggtgat gccgattatg
     2281 ttcccagcgc tctatcggat ttccaaagag cactggaatc aaacaattgt tgcacttgtc
     2341 tataatgtac tcaaaacttt tatggaaatg aatggaaaac tgtttgatga gcttacttct
     2401 acgtacaaag gtgaacgatt gcgggaaaag caacgagaaa aggatcgtga tgctttttgg
     2461 aagaaaatgg aagctcttga attgaatcca ccggctgaag gaaaagaagt gacaccttcg
     2521 ttgtttccag agaagttgac tgattatttg aagaaggacg gtccaaatat gacgccactg
     2581 ccagttgcaa ctgcaggtgg cggtgataaa tctccatctg tagtgaagaa atcgtcaaca
     2641 ggatcagaaa cgacgacgcc ggctaaaaaa taagtcgacc aactcgctcg cccgggccga
     2701 accaggtgca gccaagctaa ttccgggcga atttcttatg atttatgatt tttattatta
     2761 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
     2821 gaaaattctt gttcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagc
     2881 atgaggtcgc tcttattgac cacacctcta ccggcatgcc ggcaagtgca caaacaatac
     2941 ttaaataaat actactcagt aataacctat ttcttagcat ttttgacgaa atttgctatt
     3001 ttgttagagt cttttacacc atttgtctcc acacctccgc ttacatcaac accaataacg
     3061 ccatttaatc taagcgcatc accaacattt tctggcgtca gtccaccagc taacataaaa
     3121 tgtaagcttt cggggctctc ttgccttcca acccagtcag aaatcgagtt ccaatccaaa
     3181 agttcacctg tcccacctgc ttctgaatca aacaagggaa taaacgaatg aggtttctgt
     3241 gaagctgcac tgagtagtat gttgcagtct tttggaaata cgagtctttt aataactggc
     3301 aaaccgagga actcttggta ttcttgccac gactcatctc catgcagttg gacgatatca
     3361 atgccgtaat cattgaccag agccaaaaca tcctccttag gttgattacg aaacacgcca
     3421 accaagtatt tcggagtgcc tgaactattt ttatatgctt ttacaagact tgaaattttc
     3481 cttgcaataa ccgggtcaat tgttctcttt ctattgggca cacatataat acccagcaag
     3541 tcagcatcgg aatctagagc acattctgcg gcctctgtgc tctgcaagcc gcaaactttc
     3601 accaatggac cagaactacc tgtgaaatta ataacagaca tactccaagc tgcctttgtg
     3661 tgcttaatca cgtatactca cgtgctcaat agtcaccaat gccctccctc ttggccctct
     3721 ccttttcttt tttcgaccga attaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     3781 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3841 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3901 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3961 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4021 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4081 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4141 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4201 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4261 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4321 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4381 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4441 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4501 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4561 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4621 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4681 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4741 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4801 acgttaaggg attttggtca tgtggatgat ccagttcgat ttattcaaca aagccgccgt
     4861 cccgtcaagt cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga
     4921 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     4981 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     5041 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     5101 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     5161 ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat
     5221 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     5281 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     5341 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     5401 ctaatacctg gaatgctgtt ttgcctggga tcgcagtggt gagtaaccat gcatcatcag
     5461 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     5521 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     5581 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     5641 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg
     5701 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     5761 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     5821 tttgagacac aacgtggctt actaggatcg atccgacatg acattaacct ataaaaatag
     5881 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     5941 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     6001 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     6061 agagcagatt gtactgagag tgcaccataa cgcatttaag cataaacacg cactatgccg
     6121 ttcttctcat gtatatatat atacaggcaa cacgcagata taggtgcgac gtgaacagtg
     6181 agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg aaacgctttg
     6241 aagttcctat tccgaagttc ctattctcta gctagaaagt ataggaactt cagagcgctt
     6301 ttgaaaacca aaagcgctct gaagacgcac tttcaaaaaa ccaaaaacgc accggactgt
     6361 aacgagctac taaaatattg cgaataccgc ttccacaaac attgctcaaa agtatctctt
     6421 tgctatatat ctctgtgcta tatccctata taacctaccc atccaccttt cgctccttga
     6481 acttgcatct aaactcgacc tctacatttt ttatgtttat ctctagtatt actctttaga
     6541 caaaaaaatt gtagtaagaa ctattcatag agtgaatcga aaacaatacg aaaatgtaaa
     6601 catttcctat acgtagtata tagagacaaa atagaagaaa ccgttcataa ttttctgacc
     6661 aatgaagaat catcaacgct atcactttct gttcacaaag tatgcgcaat ccacatcggt
     6721 atagaatata atcggggatg cctttatctt gaaaaaatgc acccgcagct tcgctagtaa
     6781 tcagtaaacg cgggaagtgg agtcaggctt tttttatgga agagaaaata gacaccaaag
     6841 tagccttctt ctaaccttaa cggacctaca gtgcaaaaag ttatcaagag actgcattat
     6901 agagcgcaca aaggagaaaa aaagtaatct aagatgcttt gttagaaaaa tagcgctctc
     6961 gggatgcatt tttgtagaac aaaaaagaag tatagattct ttgttggtaa aatagcgctc
     7021 tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg
     7081 ctctcgcgtt gcatttttgt tttacaaaaa tgaagcacag attcttcgtt ggtaaaatag
     7141 cgctttcgcg ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat
     7201 tagcgctctc gcgttgcatt tttgttctac aaaatgaagc acagatgctt cgtt
//
