LOCUS       sequence-104104-        8302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast two-hybrid bridge vector expressing PAA-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8302)
  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 8302)
  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: pJW3.31
FEATURES             Location/Qualifiers
     source          1..8302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1847..1871)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1904..1920)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1905..1921)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      1923..2170
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1939..1957)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2188..2207)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2189..2207)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2217..2234)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2217..2233)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2226..2248)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2378..2833
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2465..2484)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2675..2696
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2964..3767)
                     /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(3768..3988)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4014..4033)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4133..4155
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4193..4211)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      4252..5594
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    4584..4631
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        5621..5725
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5726..5794,5795..6586)
                     /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(5944..5963)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6757..7345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7246..7265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7499..7516
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(7669..7674,7675..7692,7693..7699)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7707..7723
                     /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     7712..7734
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7731..7747
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7731..7747
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7766..7786
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7768..7786
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7804..8203
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             8253..8273
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    join(8295..8302,1..17)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gtacaaaaaa gcaggcttaa tgtcggttgt cgaagaagcc actgacgacg cgctctaccc
       61 gattgccgtg ctcattgacg agctgcgcaa cgaagatgtc acgttgcgac tgaacagcat
      121 tcgcaaactc tccacaatcg ctttggcgtt gggtgtggag cgcacacgca acgagctcat
      181 ccaatttttg accgacacga tttacgacga agatgaggtt ctgcttgtgc tcgccgagca
      241 actcggaaat ttcactccat tggtcggtgg tccagatcat gttcattgtc ttctccttcc
      301 acttgagaac ttggccacag tcgaggagac cgtggtccgt gacaaagccg ttgagagtct
      361 ccgcaaaatc gccgacaagc acagctcggc ctcattggag gagcatttcg tgccgatgct
      421 cagacgtttg gccactggag actggttcac atcccgtacc tcggcttgtg ggcttttctc
      481 tgttgtttat ccacgtgtca gtccagctat caagtcggaa ttgaagagca tgttccgaac
      541 cctgtgccgt gatgacaccc caatggttcg ccgtgctgct gctgcaaagc tcggagagtt
      601 cgcaaaggtg tttgagaaga ctgccgtcat cgagggtctc cactccagtc tcactgatct
      661 tcatgttgat gagcaagatt ctgttcgttt gctcactgtc gagtcagcca tcgctttcgg
      721 aactctcctc gataaggcta acaagaagaa actcatcgag ccaattctca ttgaattgtt
      781 cgatgacaag tcgtggagag ttcgttatat ggttgccgag aagcttatcg agattcagaa
      841 tgtcctcggt gaggatatgg ataccacaca tctcgttaac atgtacacga accttctcaa
      901 ggatccagag ggtgaggtta gatgtgctgc cactcagaga cttcaggaat tcgctctcaa
      961 cttgccagag gacaagcgtc agaacattat ctgcaactca ctgctcaacg ttgccaagga
     1021 attggttact gatgggaatc agctcgtcaa gtcggaactc gccggtgtca tcatgggact
     1081 tgctccactg attggcaagg aacaaactgt ctctgagctt ctcccaatct acatgcagtt
     1141 gcttaacgat caaaccccag aagttcgcct caatattatt tccagtcttg ataaggtcaa
     1201 tgaagtgatc ggagcagctc aattgtccac atccctcttg ccagccatcg ttggattggc
     1261 cgaggatgga aaatggagag ttcgactcgc aattgttcaa ttcatgccac ttcttgcttc
     1321 ccagttggga caagaattct tcgatgagaa gttgcttccg ctttgtttga actggcttac
     1381 cgaccatgtg ttctcaatcc gcgaggcatc gaccctcatc atgaaggaac tgacacagaa
     1441 gtttggagga caatgggcta gcacgaatat tgtaccaaaa atgcagaaac ttcagaagga
     1501 caccaactac ctgcagagaa tgacatgctt gttctgtttg aatacactca gtgaggcgat
     1561 gactcaggag cagattctga aggagattat gccaattgtg aaagatcttg tcgaggatga
     1621 cgtgccgaat gtcagattca acgccgcaaa gagtttgaag cgaattggaa agaacttgac
     1681 cccaagtaca ttgacgtccg aggtgaagcc actgctcgag aagctcggaa aagattcaga
     1741 ctttgacgtt cgatactttt cagaggaagc caagaactca ctcggcttgt aatcgaacat
     1801 atcgctagtc aaattggaat ctctacatct tccacttctc aatcggaccc agctttcttg
     1861 tacaaagtgg tgatgggctg caggaattcg atatcaagct tatcgatacc gtcgacctcg
     1921 agtcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac
     1981 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat
     2041 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt
     2101 gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc
     2161 tttaatttgc ggccggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     2221 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     2281 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     2341 tcccaacagt tgcgcagcct gaatggcgaa tggcgcgacg cgccctgtag cggcgcatta
     2401 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2461 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2521 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2581 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2641 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2701 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2761 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2821 acgtttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2881 gcatagggta ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat
     2941 gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc
     3001 cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt
     3061 cctcttccaa caataataat gtcagatcct gtagagacca catcatccac ggttctatac
     3121 tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa
     3181 tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct
     3241 ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc
     3301 ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc
     3361 gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct
     3421 gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg
     3481 tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc
     3541 ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa
     3601 caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc
     3661 aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca
     3721 ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt
     3781 ttcctgcagg tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct
     3841 tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc
     3901 ttctgttcgg agattaccga atcaaaaaaa tttcaaagaa accgaaatca aaaaaaagaa
     3961 taaaaaaaaa atgatgaatt gaattgaaaa gctgtggtat ggtgcactct cagtacaatc
     4021 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     4081 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4141 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     4201 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagta tgatccaata
     4261 tcaaaggaaa tgatagcatt gaaggatgag actaatccaa ttgaggagtg gcagcatata
     4321 gaacagctaa agggtagtgc tgaaggaagc atacgatacc ccgcatggaa tgggataata
     4381 tcacaggagg tactagacta cctttcatcc tacataaata gacgcatata agtacgcatt
     4441 taagcataaa cacgcactat gccgttcttc tcatgtatat atatatacag gcaacacgca
     4501 gatataggtg cgacgtgaac agtgagctgt atgtgcgcag ctcgcgttgc attttcggaa
     4561 gcgctcgttt tcggaaacgc tttgaagttc ctattccgaa gttcctattc tctagaaagt
     4621 ataggaactt cagagcgctt ttgaaaacca aaagcgctct gaagacgcac tttcaaaaaa
     4681 ccaaaaacgc accggactgt aacgagctac taaaatattg cgaataccgc ttccacaaac
     4741 attgctcaaa agtatctctt tgctatatat ctctgtgcta tatccctata taacctaccc
     4801 atccaccttt cgctccttga acttgcatct aaactcgacc tctacatttt ttatgtttat
     4861 ctctagtatt actctttaga caaaaaaatt gtagtaagaa ctattcatag agtgaatcga
     4921 aaacaatacg aaaatgtaaa catttcctat acgtagtata tagagacaaa atagaagaaa
     4981 ccgttcataa ttttctgacc aatgaagaat catcaacgct atcactttct gttcacaaag
     5041 tatgcgcaat ccacatcggt atagaatata atcggggatg cctttatctt gaaaaaatgc
     5101 acccgcagct tcgctagtaa tcagtaaacg cgggaagtgg agtcaggctt tttttatgga
     5161 agagaaaata gacaccaaag tagccttctt ctaaccttaa cggacctaca gtgcaaaaag
     5221 ttatcaagag actgcattat agagcgcaca aaggagaaaa aaagtaatct aagatgcttt
     5281 gttagaaaaa tagcgctctc gggatgcatt tttgtagaac aaaaaagaag tatagattct
     5341 ttgttggtaa aatagcgctc tcgcgttgca tttctgttct gtaaaaatgc agctcagatt
     5401 ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt tttacaaaaa tgaagcacag
     5461 attcttcgtt ggtaaaatag cgctttcgcg ttgcatttct gttctgtaaa aatgcagctc
     5521 agattctttg tttgaaaaat tagcgctctc gcgttgcatt tttgttctac aaaatgaagc
     5581 acagatgctt cgttcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5641 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5701 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5761 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5821 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5881 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5941 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6001 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6061 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6121 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6181 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6241 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6301 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6361 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6421 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6481 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6541 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6601 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6661 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6721 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6781 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6841 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6901 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6961 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7021 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7081 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7141 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7201 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7261 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7321 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7381 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7441 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     7501 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     7561 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     7621 gcgcaacgca attaatgtga gttacctcac tcattaggca ccccaggctt tacactttat
     7681 gcttccggct cctatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     7741 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
     7801 gctcatagct tcaaaatgtt tctactcctt ttttactctt ccagattttc tcggactccg
     7861 cgcatcgccg taccacttca aaacacccaa gcacagcata ctaaatttcc cctctttctt
     7921 cctctagggt gtcgttaatt acccgtacta aaggtttgga aaagaaaaaa gagaccgcct
     7981 cgtttctttt tcttcgtcga aaaaggcaat aaaaattttt atcacgtttc tttttcttga
     8041 aaattttttt ttttgatttt tttctctttc gatgacctcc cattgatatt taagttaata
     8101 aacggtcttc aatttctcaa gtttcagttt catttttctt gttctattac aacttttttt
     8161 acttcttgct cattagaaag aaagcatagc aatctaatct aagttttcta gaactagtaa
     8221 agatggataa agcggaatta attcccgagc ctccaaaaaa gaagagaaag gtcgaattgg
     8281 gtggatcccc catcacaagt tt
//
