LOCUS       sequence-182741-        8117 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express yeast Kre33 with Y2H activation domain fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8117)
  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 8117)
  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-Kre33
FEATURES             Location/Qualifiers
     source          1..8117
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     rep_origin      238..1580
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    570..617
                     /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        2057..2461
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2474..3568
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     protein_bind    4186..4219
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4263..4296
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4368..4401
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      complement(4419..4606)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     4527..4551
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(5036..5062)
                     /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"
     CDS             complement(5042..5068)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(5081..5422)
                     /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             complement(5438..5458)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(5504..5905)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     primer_bind     complement(5934..5956)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6042..6182
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6097..6116
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6197..6214)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6368..6956)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6448..6467)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7127..7918,7919..7987))
                     /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     7750..7769
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7988..8092)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gctttgaaga aaaatgcgcc ttattcaatc tttgctataa aaaatggccc aaaatctcac
       61 attggaagac atttgatgac ctcatttctt tcaatgaagg gcctaacgga gttgactaat
      121 gttgtgggaa attggagcga taagcgtgct tctgccgtgg ccaggacaac gtatactcat
      181 cagataacag caatacctga tcactacttc gcactagttt ctcggtacta tgcatatgat
      241 ccaatatcaa aggaaatgat agcattgaag gatgagacta atccaattga ggagtggcag
      301 catatagaac agctaaaggg tagtgctgaa ggaagcatac gataccccgc atggaatggg
      361 ataatatcac aggaggtact agactacctt tcatcctaca taaatagacg catataagta
      421 cgcatttaag cataaacacg cactatgccg ttcttctcat gtatatatat atacaggcaa
      481 cacgcagata taggtgcgac gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt
      541 tcggaagcgc tcgttttcgg aaacgctttg aagttcctat tccgaagttc ctattctcta
      601 gaaagtatag gaacttcaga gcgcttttga aaaccaaaag cgctctgaag acgcactttc
      661 aaaaaaccaa aaacgcaccg gactgtaacg agctactaaa atattgcgaa taccgcttcc
      721 acaaacattg ctcaaaagta tctctttgct atatatctct gtgctatatc cctatataac
      781 ctacccatcc acctttcgct ccttgaactt gcatctaaac tcgacctcta cattttttat
      841 gtttatctct agtattactc tttagacaaa aaaattgtag taagaactat tcatagagtg
      901 aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt agtatataga gacaaaatag
      961 aagaaaccgt tcataatttt ctgaccaatg aagaatcatc aacgctatca ctttctgttc
     1021 acaaagtatg cgcaatccac atcggtatag aatataatcg gggatgcctt tatcttgaaa
     1081 aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg aagtggagtc aggctttttt
     1141 tatggaagag aaaatagaca ccaaagtagc cttcttctaa ccttaacgga cctacagtgc
     1201 aaaaagttat caagagactg cattatagag cgcacaaagg agaaaaaaag taatctaaga
     1261 tgctttgtta gaaaaatagc gctctcggga tgcatttttg tagaacaaaa aagaagtata
     1321 gattctttgt tggtaaaata gcgctctcgc gttgcatttc tgttctgtaa aaatgcagct
     1381 cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgtttta caaaaatgaa
     1441 gcacagattc ttcgttggta aaatagcgct ttcgcgttgc atttctgttc tgtaaaaatg
     1501 cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg ttctacaaaa
     1561 tgaagcacag atgcttcgtt aacaaagata tgctattgaa gtgcaagatg gaaacgcaga
     1621 aaatgaaccg gggatgcgac gtgcaagatt acctatgcaa tagatgcaat agtttctcca
     1681 ggaaccgaaa tacatacatt gtcttccgta aagcgctaga ctatatatta ttatacaggt
     1741 tcaaatatac tatctgtttc agggaaaact cccaggttcg gatgttcaaa attcaatgat
     1801 gggtaacaag tacgatcgta aatctgtaaa acagtttgtc ggatattagg ctgtatctcc
     1861 tcaaagcgta ttcgaatatc attgagaagc tgcagcgtca catcggataa taatgatggc
     1921 agccattgta gaagtgcctt ttgcatttct agtctctttc tcggtctagc tagttttact
     1981 acatcgcgaa gatagaatct tagatcacac tgcctttgct gagctggatc aatagagtaa
     2041 caaaagagtg gtaaggaact gtgggaatac tcaggtatcg taagatgcaa gagttcgaat
     2101 ctcttagcaa ccattatttt tttcctcaac ataacgagaa cacacagggg cgctatcgca
     2161 cagaatcaaa ttcgatgact ggaaattttt tgttaatttc agaggtcgcc tgacgcatat
     2221 acctttttca actgaaaaat tgggagaaaa aggaaaggtg agaggccgga accggctttt
     2281 catatagaat agagaagcgt tcatgactaa atgcttgcat cacaatactt gaagttgaca
     2341 atattattta aggacctatt gttttttcca ataggtggtt agcaatcgtc ttactttcta
     2401 acttttctta ccttttacat ttcagcaata tatatatata tttcaaggat ataccattct
     2461 aatgtctgcc cctatgtctg cccctaagaa gatcgtcgtt ttgccaggtg accacgttgg
     2521 tcaagaaatc acagccgaag ccattaaggt tcttaaagct atttctgatg ttcgttccaa
     2581 tgtcaagttc gatttcgaaa atcatttaat tggtggtgct gctatcgatg ctacaggtgt
     2641 cccacttcca gatgaggcgc tggaagcctc caagaaggtt gatgccgttt tgttaggtgc
     2701 tgtgggtggt cctaaatggg gtaccggtag tgttagacct gaacaaggtt tactaaaaat
     2761 ccgtaaagaa cttcaattgt acgccaactt aagaccatgt aactttgcat ccgactctct
     2821 tttagactta tctccaatca agccacaatt tgctaaaggt actgacttcg ttgttgtcag
     2881 agaattagtg ggaggtattt actttggtaa gagaaaggaa gacgatggtg atggtgtcgc
     2941 ttgggatagt gaacaataca ccgttccaga agtgcaaaga atcacaagaa tggccgcttt
     3001 catggcccta caacatgagc caccattgcc tatttggtcc ttggataaag ctaatgtttt
     3061 ggcctcttca agattatgga gaaaaactgt ggaggaaacc atcaagaacg aatttcctac
     3121 attgaaggtt caacatcaat tgattgattc tgccgccatg atcctagtta agaacccaac
     3181 ccacctaaat ggtattataa tcaccagcaa catgtttggt gatatcatct ccgatgaagc
     3241 ctccgttatc ccaggttcct tgggtttgtt gccatctgcg tccttggcct ctttgccaga
     3301 caagaacacc gcatttggtt tgtacgaacc atgccacggt tctgctccag atttgccaaa
     3361 gaataaggtt gaccctatcg ccactatctt gtctgctgca atgatgttga aattgtcatt
     3421 gaacttgcct gaagaaggta aggccattga agatgcagtt aaaaaggttt tggatgcagg
     3481 tatcagaact ggtgatttag gtggttccaa cagtaccacc gaagtcggtg atgctgtcgc
     3541 cgaagaagtt aagaaaatcc ttgcttaaaa agattctctt tttttatgat atttgtacat
     3601 aaactttata aatgaaattc ataatagaaa cgacacgaaa ttacaaaatg gaatatgttc
     3661 atagggtaga cgaaactata tacgcaatct acatacattt atcaagaagg agaaaaagga
     3721 ggatagtaaa ggaatacagg taagcaaatt gatactaatg gctcaacgtg ataaggaaaa
     3781 agaattgcac tttaacatta atattgacaa ggaggagggc accacacaaa aagttaggtg
     3841 taacagaaaa tcatgaaact acgattccta atttgatatt ggaggatttt ctctaaaaaa
     3901 aaaaaaatac aacaaataaa aaacactcaa tgacctgacc atttgatgga gtttaagtca
     3961 ataccttctt gaaccatttc ccataatggt gaaagttccc tcaagaattt tactctgtca
     4021 gaaacggcct taacgacgta gtcgaccaat tccgatcata tcaataaccc ttaatataac
     4081 ttcgtataat gtatgctatc gaagttatta ggtctgaaga ggagtttacg tccagccaag
     4141 ctagcctggc tgcaggtcga gcggccgcga tccggaaccc ttaatataac ttcgtataat
     4201 gtatgctata cgaagttatt aggtccctcg accaattccg atcatattca ataaccctta
     4261 atataacttc gtataatgta tgctatacga agttattagg tctgaagagg agtttacgtc
     4321 cgctagcttg gctgcaggtc gagcggccgc gatccggaac ccttaatata acttcgtata
     4381 atgtatgcta tacgaagtta ttaggtccct cgcacatgcc ggtagaggtg tggtcaataa
     4441 gagcgacctc atgctatacc tgagaaagca acctgaccta caggaaagag ttactcaaga
     4501 acaagaattt tcgttttaaa acctaagagt cactttaaaa tttgtataca cttatttttt
     4561 ttataactta tttaataata aaaatcataa atcataagaa attcgcttat ttagaagtgt
     4621 caacaacgta tctaccaacg atttgaccct tttccatctt ttcgtaaatt tctggcaagg
     4681 tagacaagcc gacaaccttg attggagact tgaccaaacc tctggcgaag aagtccaaag
     4741 cttctgaata agccctcgta atatattttc atgaagaatt taggtccaaa aaaaagatgg
     4801 gcattaattc tagtcattta aaaaattcta tagatcagag gttacatggc caagattgaa
     4861 acttagagga gtatagttac ataaaagaag gcaaaacgat gtataaatga aagaaattga
     4921 gatggtgcac gatgcacagt tgaagtgaac ttgcggggtt tttcagtatc tacgattcat
     4981 agatctctcg agctcgaatt cggatccccg gggcctccat ggccatatga ccacccaagc
     5041 tagcgtaatc tggaacatcg tatgggtaag ccatacagat ctcttttttt gggtttggtg
     5101 gggtatcttc atcatcgaat agatagttat atacatcatc cattgtagtg gtattaaaca
     5161 tccctgtagt gattccaaac gcgttatacg cagtttggtc cgtccaacca ggtgacagtg
     5221 gttttgaatt attaccatca tcaattttac tagccgtgat ttcattattc atgaagttat
     5281 catgaacgtt agaggaggca attggttgtg aaagcgcttg agaatttgtt tgagttgtta
     5341 tgaggttcgg accgttgcta ctgttagtga aagtgaagga caatgagcta tcagcaatat
     5401 tcccactttg attaaaattg gcggcggtac ccaattcgac ctttctcttc ttttttggag
     5461 gctcgggaat taattccgct ttatccatct ttgcaaagct tggagttgat tgtatgcttg
     5521 gtatagcttg aaatattgtg cagaaaaaga aacaaggaag aaagggaacg agaacaatga
     5581 cgaggaaaca aaagattaat aattgcaggt ctatttatac ttgatagcaa agcggcaaac
     5641 tttttttatt tcaaattcaa gtaactggaa ggaaggccgt ataccgttgc tcattagaga
     5701 gtagtgtgcg tgaatgaagg aaggaaaaag tttcgtgtgt tcgaagatac ccctcatcag
     5761 ctctggaaca acgacatctg ttggtgctgt ctttgtcgtt aattttttcc tttagtgtct
     5821 tccatcattt tttttgtcat tgcggatatg gtgagacaac aacgggggag agagaaaaga
     5881 aaaaaaaaga aaagaagttg ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     5941 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     6001 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     6061 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga
     6121 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     6181 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6241 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6301 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6361 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6421 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6481 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6541 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6601 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6661 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6721 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6781 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6841 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6901 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6961 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7021 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7081 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7141 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7201 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7261 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7321 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7381 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7441 ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7501 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7561 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7621 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7681 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7741 caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7801 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7861 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7921 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7981 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8041 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8101 cccgaaaagt gccacct
//
