LOCUS       sequence-191952-        9407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express yeast Utp15 with Y2H activation domain fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9407)
  AUTHORS   Vincent NG, Charette JM, Baserga SJ
  TITLE     The SSU processome interactome in Saccharomyces cerevisiae reveals 
            potential new protein subcomplexes.
  JOURNAL   RNA. 2017 Oct 20. pii: rna.062927.117. doi: 10.1261/rna.062927.117.
  PUBMED    29054886
REFERENCE   2  (bases 1 to 9407)
  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: "RNA. 2017 
            Oct 20. pii: rna.062927.117. doi: 10.1261/rna.062927.117."
COMMENT     Sequence Label: pACT2-Utp15
FEATURES             Location/Qualifiers
     source          1..9407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..397
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             443..463
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             479..820
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     CDS             833..859
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             839..865
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     protein_bind    899..923
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2469..2493)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      2882..3069
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2937..2961)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    complement(3090..3123)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3632..4726)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4727..5132)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      5609..6951
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(6572..6619)
                     /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        7216..7320
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7321..7389,7390..8181)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7539..7558)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8352..8940
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8841..8860
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9094..9111
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    9126..9266
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(9192..9211)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9352..9374
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 caacttcttt tctttttttt tcttttctct ctcccccgtt gttgtctcac catatccgca
       61 atgacaaaaa aatgatggaa gacactaaag gaaaaaatta acgacaaaga cagcaccaac
      121 agatgtcgtt gttccagagc tgatgagggg tatctcgaag cacacgaaac tttttccttc
      181 cttcattcac gcacactact ctctaatgag caacggtata cggccttcct tccagttact
      241 tgaatttgaa ataaaaaaaa gtttgctgtc ttgctatcaa gtataaatag acctgcaatt
      301 attaatcttt tgtttcctcg tcattgttct cgttcccttt cttccttgtt tctttttctg
      361 cacaatattt caagctatac caagcataca atcaactcca agctttgcaa agatggataa
      421 agcggaatta attcccgagc ctccaaaaaa gaagagaaag gtcgaattgg gtaccgccgc
      481 caattttaat caaagtggga atattgctga tagctcattg tccttcactt tcactaacag
      541 tagcaacggt ccgaacctca taacaactca aacaaattct caagcgcttt cacaaccaat
      601 tgcctcctct aacgttcatg ataacttcat gaataatgaa atcacggcta gtaaaattga
      661 tgatggtaat aattcaaaac cactgtcacc tggttggacg gaccaaactg cgtataacgc
      721 gtttggaatc actacaggga tgtttaatac cactacaatg gatgatgtat ataactatct
      781 attcgatgat gaagataccc caccaaaccc aaaaaaagag atctctatgg cttacccata
      841 cgatgttcca gattacgcta gcttgggtgg tcatatggcc atggaggccc cagctttcac
      901 aagtttgtac aaaaaagcag gctacaaaat gtcgactgct aggcctagaa taatcacttc
      961 gaaggctcca ttattgcctc aacagactac cccggagcag cgttactggc gtcaatacac
     1021 ttccgcacaa ctggttaaag aacataatag cgttactcac atatcattca atcctcaaca
     1081 tcctcatgat tttgcagtaa cttcctcaac cagagttcaa atattttctt caagaactag
     1141 gcaagtgatc aagacctttt caagatttaa agatgttgtc tattctgcat cctttaggag
     1201 tgatggtaaa ttgctttgtg ccggtgatgc taccggttta gtgtctgtat acgacagtta
     1261 taatcccagg actatccttt tatctataaa cgcttccaca catcccacac atgttacgaa
     1321 attccatacg caagataaca aaattctggc tacggcgagt gacgatagag tgacaagact
     1381 ttgggatata tccaacgcat atgagccaca attagagctc accggagcta ctgactatgt
     1441 tcgtacctta tcatttatcc cagcagcccc acatttggtt gcaacaggct catacgacgg
     1501 tttgatcaga ttatatgaca caagatctag cggatccacg ccaatttatt ctctgaatca
     1561 tgaccaacct gttgaaaatg ttattgcagt atctccaacc caaattgttt cttgcggtgg
     1621 caataatttc aaagtgtggg acttgaccag taacaagaaa ttatacgaac gtggcaattt
     1681 caataaggct gtgacatgct tggactatgt tgaaaatttt gattctccaa tgcaatcggc
     1741 gttgatcgct tcttccttag atggtcatgt caaagtcttt gatcctttag acaatttcca
     1801 agtcaaattt ggctggaaat tttcgggtcc cgtactaagt tgtgctgtct cccctagtac
     1861 cgcacaaggg aataggcatt tagtagcagg tttatcatca ggactactag ccattagaac
     1921 taaaaagaaa gagaaacgtt cttccgataa agaaaacgca cccgccagtt ttaataagaa
     1981 tgcgaaaagt aataacttcc agagaatgat gcgaggctcg gaataccaag gtgatcaaga
     2041 gcatataatc cacaatgata aggtaagatc tcaacgtcgt atgcgtgcat tcgaaaggaa
     2101 cataaatcag tttaaatgga gcgaagcact cgacaatgcc tttgtaccag gtatggccaa
     2161 agagttgaca cttactgttc ttcaagaact acgcaaacgt ggcaaagttc gtgtagcgct
     2221 atatggtaga gatgaatcta cattagagcc gctactgaat tggtgcttaa aaggtattga
     2281 agatgtgaga tcagcatcta ttgtagcgga ttgggtcgcc gtggttttag aattatatgg
     2341 gaatacgtta gaaagctctc cagttcttca agaattgatg atagatttga aaactaaagt
     2401 tagacatgaa attcataagt ctaaagaagc tcaaagaatt gaaggtatgc ttcaactttt
     2461 aacgagtaac ccagctttct tgtacaaagt ggtgagtcga gagatctatg aatcgtagat
     2521 actgaaaaac cccgcaagtt cacttcaact gtgcatcgtg caccatctca atttctttca
     2581 tttatacatc gttttgcctt cttttatgta actatactcc tctaagtttc aatcttggcc
     2641 atgtaacctc tgatctatag aattttttaa atgactagaa ttaatgccca tctttttttt
     2701 ggacctaaat tcttcatgaa aatatattac gagggcttat tcagaagctt tggacttctt
     2761 cgccagaggt ttggtcaagt ctccaatcaa ggttgtcggc ttgtctacct tgccagaaat
     2821 ttacgaaaag atggaaaagg gtcaaatcgt tggtagatac gttgttgaca cttctaaata
     2881 agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt
     2941 atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc
     3001 tttcctgtag gtcaggttgc tttctcaggt atagcatgag gtcgctctta ttgaccacac
     3061 ctctaccggc atgccggtcg agggacctaa taacttcgta tagcatacat tatacgaagt
     3121 tatattaagg gttattgaat atgatcggaa ttggtcgact acgtcgttaa ggccgtttct
     3181 gacagagtaa aattcttgag ggaactttca ccattatggg aaatggttca agaaggtatt
     3241 gacttaaact ccatcaaatg gtcaggtcat tgagtgtttt ttatttgttg tatttttttt
     3301 tttttagaga aaatcctcca atatataaat taggaatcat agtttcatga ttttctgtta
     3361 cacctaactt tttgtgtggt gccctcctcc ttgtcaatat taatgttaaa gtgcaattct
     3421 ttttccttat cacgttgagc cattagtatc aatttgctta cctgtattcc tttacatcct
     3481 cctttttctc cttcttgata aatgtatgta gattgcgtat atagtttcgt ctaccctatg
     3541 aacatattcc attttgtaat ttcgtgtcgt ttctattatg aatttcattt ataaagttta
     3601 tgtacaaata tcataaaaaa agagaatctt tttaagcaag gattttctta acttcttcgg
     3661 cgacagcatc accgacttcg gtggtactgt tggaaccacc taaatcacca gttctgatac
     3721 ctgcatccaa aaccttttta actgcatctt caatggcctt accttcttca ggcaagttca
     3781 atgacaattt caacatcatt gcagcagaca agatagtggc gatagggttg accttattct
     3841 ttggcaaatc tggagcagaa ccgtggcatg gttcgtacaa accaaatgcg gtgttcttgt
     3901 ctggcaaaga ggccaaggac gcagatggca acaaacccaa ggaacctggg ataacggagg
     3961 cttcatcgga gatgatatca ccaaacatgt tgctggtgat tataatacca tttaggtggg
     4021 ttgggttctt aactaggatc atggcggcag aatcaatcaa ttgatgttga accttcaatg
     4081 taggaaattc gttcttgatg gtttcctcca cagtttttct ccataatctt gaagaggcca
     4141 aaacattagc tttatccaag gaccaaatag gcaatggtgg ctcatgttgt agggccatga
     4201 aagcggccat tcttgtgatt ctttgcactt ctggaacggt gtattgttca ctatcccaag
     4261 cgacaccatc accatcgtct tcctttctct taccaaagta aatacctccc actaattctc
     4321 tgacaacaac gaagtcagta cctttagcaa attgtggctt gattggagat aagtctaaaa
     4381 gagagtcgga tgcaaagtta catggtctta agttggcgta caattgaagt tctttacgga
     4441 tttttagtaa accttgttca ggtctaacac tacctgtacc ccatttagga ccacccacag
     4501 cacctaacaa aacggcatca gccttcttgg aggcttccag cgcctcatct ggaagtggaa
     4561 cacctgtagc atcgatagca gcaccaccaa ttaaatgatt ttcgaaatcg aacttgacat
     4621 tggaacgaac atcagaaata gctttaagaa ccttaatggc ttcggctgtg atttcttgac
     4681 caacgtggtc acctggcaaa acgacgatct tcttaggggc agacattaga atggtatatc
     4741 cttgaaatat atatatatat tgctgaaatg taaaaggtaa gaaaagttag aaagtaagac
     4801 gattgctaac cacctattgg aaaaaacaat aggtccttaa ataatattgt caacttcaag
     4861 tattgtgatg caagcattta gtcatgaacg cttctctatt ctatatgaaa agccggttcc
     4921 ggcgctctca cctttccttt ttctcccaat ttttcagttg aaaaaggtat atgcgtcagg
     4981 cgacctctga aattaacaaa aaatttccag tcatcgaatt tgattctgtg cgatagcgcc
     5041 cctgtgtgtt ctcgttatgt tgaggaaaaa aataatggtt gctaagagat tcgaactctt
     5101 gcatcttacg atacctgagt attcccacag ttccttacca ctcttttgtt actctattga
     5161 tccagctcag caaaggcagt gtgatctaag attctatctt cgcgatgtag taaaactagc
     5221 tagaccgaga aagagactag aaatgcaaaa ggcacttcta caatggctgc catcattatt
     5281 atccgatgtg acgctgcagc ttctcaatga tattcgaata cgctttgagg agatacagcc
     5341 taatatccga caaactgttt tacagattta cgatcgtact tgttacccat cattgaattt
     5401 tgaacatccg aacctgggag ttttccctga aacagatagt atatttgaac ctgtataata
     5461 atatatagtc tagcgcttta cggaagacaa tgtatgtatt tcggttcctg gagaaactat
     5521 tgcatctatt gcataggtaa tcttgcacgt cgcatccccg gttcattttc tgcgtttcca
     5581 tcttgcactt caatagcata tctttgttaa cgaagcatct gtgcttcatt ttgtagaaca
     5641 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     5701 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta
     5761 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     5821 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
     5881 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact
     5941 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
     6001 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
     6061 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
     6121 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
     6181 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
     6241 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
     6301 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
     6361 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
     6421 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
     6481 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtct
     6541 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gagaatagga
     6601 acttcggaat aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa atgcaacgcg
     6661 agctgcgcac atacagctca ctgttcacgt cgcacctata tctgcgtgtt gcctgtatat
     6721 atatatacat gagaagaacg gcatagtgcg tgtttatgct taaatgcgta cttatatgcg
     6781 tctatttatg taggatgaaa ggtagtctag tacctcctgt gatattatcc cattccatgc
     6841 ggggtatcgt atgcttcctt cagcactacc ctttagctgt tctatatgct gccactcctc
     6901 aattggatta gtctcatcct tcaatgctat catttccttt gatattggat catatgcata
     6961 gtaccgagaa actagtgcga agtagtgatc aggtattgct gttatctgat gagtatacgt
     7021 tgtcctggcc acggcagaag cacgcttatc gctccaattt cccacaacat tagtcaactc
     7081 cgttaggccc ttcattgaaa gaaatgaggt catcaaatgt cttccaatgt gagattttgg
     7141 gccatttttt atagcaaaga ttgaataagg cgcatttttc ttcaaagctc aggtggcact
     7201 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     7261 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     7321 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     7381 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     7441 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     7501 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     7561 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     7621 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     7681 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     7741 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     7801 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     7861 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     7921 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7981 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     8041 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     8101 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     8161 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     8221 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     8281 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     8341 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     8401 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     8461 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     8521 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     8581 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     8641 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     8701 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     8761 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     8821 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     8881 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8941 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     9001 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     9061 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     9121 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     9181 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     9241 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     9301 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     9361 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcag
//
