LOCUS       sequence-192202-        8576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express yeast Mrt4 with Y2H activation domain fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8576)
  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 8576)
  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-Mrt4
FEATURES             Location/Qualifiers
     source          1..8576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..141
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(67..86)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     227..249
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        278..679
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             725..745
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             761..1102
                     /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             1115..1141
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1121..1147
                     /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    1181..1205
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1920..1944)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      2333..2520
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2388..2412)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    complement(2541..2574)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3083..4177)
                     /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(4178..4583)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      5060..6402
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(6023..6070)
                     /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        6667..6771
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6772..6840,6841..7632)
                     /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(6990..7009)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7803..8391
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8292..8311
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8545..8562
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
       61 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
      121 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
      181 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
      241 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcaacttct tttctttttt
      301 tttcttttct ctctcccccg ttgttgtctc accatatccg caatgacaaa aaaatgatgg
      361 aagacactaa aggaaaaaat taacgacaaa gacagcacca acagatgtcg ttgttccaga
      421 gctgatgagg ggtatctcga agcacacgaa actttttcct tccttcattc acgcacacta
      481 ctctctaatg agcaacggta tacggccttc cttccagtta cttgaatttg aaataaaaaa
      541 aagtttgctg tcttgctatc aagtataaat agacctgcaa ttattaatct tttgtttcct
      601 cgtcattgtt ctcgttccct ttcttccttg tttctttttc tgcacaatat ttcaagctat
      661 accaagcata caatcaactc caagctttgc aaagatggat aaagcggaat taattcccga
      721 gcctccaaaa aagaagagaa aggtcgaatt gggtaccgcc gccaatttta atcaaagtgg
      781 gaatattgct gatagctcat tgtccttcac tttcactaac agtagcaacg gtccgaacct
      841 cataacaact caaacaaatt ctcaagcgct ttcacaacca attgcctcct ctaacgttca
      901 tgataacttc atgaataatg aaatcacggc tagtaaaatt gatgatggta ataattcaaa
      961 accactgtca cctggttgga cggaccaaac tgcgtataac gcgtttggaa tcactacagg
     1021 gatgtttaat accactacaa tggatgatgt atataactat ctattcgatg atgaagatac
     1081 cccaccaaac ccaaaaaaag agatctctat ggcttaccca tacgatgttc cagattacgc
     1141 tagcttgggt ggtcatatgg ccatggaggc cccagctttc acaagtttgt acaaaaaagc
     1201 aggctacaaa atgccaagat caaaacgttc caagctagtc actttagcac aaaccgataa
     1261 aaagggcaga gaaaataaag aaagaatttt cgatgaagtg agggaagcct tagatactta
     1321 tagatacgtc tgggtcctac atcttgacga cgtaagaact ccagttttac aagaaatcag
     1381 aacatcttgg gcaggctcta agttaattat gggtaagagg aaagttttac aaaaagcatt
     1441 gggagaaaag agggaagaag aatacaaaga aaatctgtat caattgagta agctttgtag
     1501 cggtgtcact ggtttattgt tcactgatga ggatgtcaac actgtcaagg aatactttaa
     1561 gtcatacgtt cgttcagact attcaagacc gaacacgaaa gcaccactaa catttacaat
     1621 tcctgagggc attgtctact cacgtggtgg tcaaattcca gctgaggaag atgttccaat
     1681 gattcattct ttagagccaa ctatgagaaa caaatttgaa attccaacta aaatcaaagc
     1741 tggtaagatc accattgaca gtccatattt agtttgcact gaaggagaaa aattagatgt
     1801 tcgtcaagct ttaatactga aacaattcgg tatcgctgct tctgaattca aagtcaaggt
     1861 gtcggcctac tatgacaacg atagctccac tgttgaaagc actaacatca acatggaaaa
     1921 cccagctttc ttgtacaaag tggtgagtcg agagatctat gaatcgtaga tactgaaaaa
     1981 ccccgcaagt tcacttcaac tgtgcatcgt gcaccatctc aatttctttc atttatacat
     2041 cgttttgcct tcttttatgt aactatactc ctctaagttt caatcttggc catgtaacct
     2101 ctgatctata gaatttttta aatgactaga attaatgccc atcttttttt tggacctaaa
     2161 ttcttcatga aaatatatta cgagggctta ttcagaagct ttggacttct tcgccagagg
     2221 tttggtcaag tctccaatca aggttgtcgg cttgtctacc ttgccagaaa tttacgaaaa
     2281 gatggaaaag ggtcaaatcg ttggtagata cgttgttgac acttctaaat aagcgaattt
     2341 cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt
     2401 ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta
     2461 ggtcaggttg ctttctcagg tatagcatga ggtcgctctt attgaccaca cctctaccgg
     2521 catgccggtc gagggaccta ataacttcgt atagcataca ttatacgaag ttatattaag
     2581 ggttattgaa tatgatcgga attggtcgac tacgtcgtta aggccgtttc tgacagagta
     2641 aaattcttga gggaactttc accattatgg gaaatggttc aagaaggtat tgacttaaac
     2701 tccatcaaat ggtcaggtca ttgagtgttt tttatttgtt gtattttttt ttttttagag
     2761 aaaatcctcc aatatataaa ttaggaatca tagtttcatg attttctgtt acacctaact
     2821 ttttgtgtgg tgccctcctc cttgtcaata ttaatgttaa agtgcaattc tttttcctta
     2881 tcacgttgag ccattagtat caatttgctt acctgtattc ctttacatcc tcctttttct
     2941 ccttcttgat aaatgtatgt agattgcgta tatagtttcg tctaccctat gaacatattc
     3001 cattttgtaa tttcgtgtcg tttctattat gaatttcatt tataaagttt atgtacaaat
     3061 atcataaaaa aagagaatct ttttaagcaa ggattttctt aacttcttcg gcgacagcat
     3121 caccgacttc ggtggtactg ttggaaccac ctaaatcacc agttctgata cctgcatcca
     3181 aaaccttttt aactgcatct tcaatggcct taccttcttc aggcaagttc aatgacaatt
     3241 tcaacatcat tgcagcagac aagatagtgg cgatagggtt gaccttattc tttggcaaat
     3301 ctggagcaga accgtggcat ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag
     3361 aggccaagga cgcagatggc aacaaaccca aggaacctgg gataacggag gcttcatcgg
     3421 agatgatatc accaaacatg ttgctggtga ttataatacc atttaggtgg gttgggttct
     3481 taactaggat catggcggca gaatcaatca attgatgttg aaccttcaat gtaggaaatt
     3541 cgttcttgat ggtttcctcc acagtttttc tccataatct tgaagaggcc aaaacattag
     3601 ctttatccaa ggaccaaata ggcaatggtg gctcatgttg tagggccatg aaagcggcca
     3661 ttcttgtgat tctttgcact tctggaacgg tgtattgttc actatcccaa gcgacaccat
     3721 caccatcgtc ttcctttctc ttaccaaagt aaatacctcc cactaattct ctgacaacaa
     3781 cgaagtcagt acctttagca aattgtggct tgattggaga taagtctaaa agagagtcgg
     3841 atgcaaagtt acatggtctt aagttggcgt acaattgaag ttctttacgg atttttagta
     3901 aaccttgttc aggtctaaca ctacctgtac cccatttagg accacccaca gcacctaaca
     3961 aaacggcatc agccttcttg gaggcttcca gcgcctcatc tggaagtgga acacctgtag
     4021 catcgatagc agcaccacca attaaatgat tttcgaaatc gaacttgaca ttggaacgaa
     4081 catcagaaat agctttaaga accttaatgg cttcggctgt gatttcttga ccaacgtggt
     4141 cacctggcaa aacgacgatc ttcttagggg cagacattag aatggtatat ccttgaaata
     4201 tatatatata ttgctgaaat gtaaaaggta agaaaagtta gaaagtaaga cgattgctaa
     4261 ccacctattg gaaaaaacaa taggtcctta aataatattg tcaacttcaa gtattgtgat
     4321 gcaagcattt agtcatgaac gcttctctat tctatatgaa aagccggttc cggcgctctc
     4381 acctttcctt tttctcccaa tttttcagtt gaaaaaggta tatgcgtcag gcgacctctg
     4441 aaattaacaa aaaatttcca gtcatcgaat ttgattctgt gcgatagcgc ccctgtgtgt
     4501 tctcgttatg ttgaggaaaa aaataatggt tgctaagaga ttcgaactct tgcatcttac
     4561 gatacctgag tattcccaca gttccttacc actcttttgt tactctattg atccagctca
     4621 gcaaaggcag tgtgatctaa gattctatct tcgcgatgta gtaaaactag ctagaccgag
     4681 aaagagacta gaaatgcaaa aggcacttct acaatggctg ccatcattat tatccgatgt
     4741 gacgctgcag cttctcaatg atattcgaat acgctttgag gagatacagc ctaatatccg
     4801 acaaactgtt ttacagattt acgatcgtac ttgttaccca tcattgaatt ttgaacatcc
     4861 gaacctggga gttttccctg aaacagatag tatatttgaa cctgtataat aatatatagt
     4921 ctagcgcttt acggaagaca atgtatgtat ttcggttcct ggagaaacta ttgcatctat
     4981 tgcataggta atcttgcacg tcgcatcccc ggttcatttt ctgcgtttcc atcttgcact
     5041 tcaatagcat atctttgtta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa
     5101 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     5161 caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa
     5221 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     5281 aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac
     5341 aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc
     5401 ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt
     5461 tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc
     5521 ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc
     5581 gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg
     5641 atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta
     5701 cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac
     5761 tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag
     5821 tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca
     5881 gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt
     5941 ttagtagctc gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc ttcagagcgc
     6001 ttttggtttt caaaagcgct ctgaagttcc tatactttct agagaatagg aacttcggaa
     6061 taggaacttc aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca
     6121 catacagctc actgttcacg tcgcacctat atctgcgtgt tgcctgtata tatatataca
     6181 tgagaagaac ggcatagtgc gtgtttatgc ttaaatgcgt acttatatgc gtctatttat
     6241 gtaggatgaa aggtagtcta gtacctcctg tgatattatc ccattccatg cggggtatcg
     6301 tatgcttcct tcagcactac cctttagctg ttctatatgc tgccactcct caattggatt
     6361 agtctcatcc ttcaatgcta tcatttcctt tgatattgga tcatatgcat agtaccgaga
     6421 aactagtgcg aagtagtgat caggtattgc tgttatctga tgagtatacg ttgtcctggc
     6481 cacggcagaa gcacgcttat cgctccaatt tcccacaaca ttagtcaact ccgttaggcc
     6541 cttcattgaa agaaatgagg tcatcaaatg tcttccaatg tgagattttg ggccattttt
     6601 tatagcaaag attgaataag gcgcattttt cttcaaagct caggtggcac ttttcgggga
     6661 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6721 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6781 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6841 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6901 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6961 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     7021 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     7081 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     7141 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     7201 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     7261 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     7321 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     7381 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     7441 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     7501 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     7561 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     7621 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     7681 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     7741 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7801 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7861 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7921 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     7981 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     8041 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     8101 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     8161 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     8221 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     8281 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     8341 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     8401 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     8461 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     8521 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcg
//
