LOCUS       sequence-387526-        9415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expesses S.cerevisiae eIF4G1 amino acids 1-341 as Gal4 activation 
            domain fusion..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9415)
  AUTHORS   Melamed D, Young DL, Miller CR, Fields S
  TITLE     Combining natural sequence variation with high throughput mutational
            data to reveal protein interaction sites.
  JOURNAL   PLoS Genet. 2015 Feb 11;11(2):e1004918. doi: 
            10.1371/journal.pgen.1004918. eCollection 2015 Feb.
  PUBMED    25671604
REFERENCE   2  (bases 1 to 9415)
  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: "PLoS 
            Genet."; date: "2015-02-11"; volume: "11(2):e1004918. doi"; pages: "
            10.1371/journal.pgen.1004918. eCollection 2015 Feb"
COMMENT     Sequence Label: pOAD eIF4G1 (1-341)
FEATURES             Location/Qualifiers
     source          1..9415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      393..580
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(448..472)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    1410..2572
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      1735..2572
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(2934..4028)
                     /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(4029..4434)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(4454..4472)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4540..4644
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4645..4713,4714..5505)
                     /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(4863..4882)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5676..6264
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6165..6184
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6418..6435
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        7253..7957
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             8003..8023
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8039..8380
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
ORIGIN
        1 gtcgacctgc agagatctat gaatcgtaga tactgaaaaa ccccgcaagt tcacttcaac
       61 tgtgcatcgt gcaccatctc aatttctttc atttatacat cgttttgcct tcttttatgt
      121 aactatactc ctctaagttt caatcttggc catgtaacct ctgatctata gaatttttta
      181 aatgactaga attaatgccc atcttttttt tggacctaaa ttcttcatga aaatatatta
      241 cgagggctta ttcagaagct ttggacttct tcgccagagg tttggtcaag tctccaatca
      301 aggttgtcgg cttgtctacc ttgccagaaa tttacgaaaa gatggaaaag ggtcaaatcg
      361 ttggtagata cgttgttgac acttctaaat aagcgaattt cttatgattt atgattttta
      421 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
      481 taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
      541 tatagcatga ggtcgctctt attgaccaca cctctaccgg catgccgagc aaatgcctgc
      601 aaatcgctcc ccatttcacc caattgtaga tatgctaact ccagcaatga gttgatgaat
      661 ctcggtgtgt attttatgtc ctcagaggac aacacctgtt gtaatcgttc ttccacacgg
      721 atcctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
      781 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
      841 ccgcataatc gctgggccat tctcatgaag aatatcttga atttattgtc atattactag
      901 ttggtgtgga agtcctaata tcggtgatca atatagtggt tgacatgctg gctagtcaac
      961 attgagcctt ttgatcatgc aaatatatta cggtatttta caatcaaata tcaaacttaa
     1021 ctattgactt tataacttat ttaggtggta acattcttat aaaaaagaaa aaaattactg
     1081 caaaacagta ctagctttta acttgtatcc taggttatct atgctgtctc accatagaga
     1141 atattaccta tttcagaatg tatgtccatg attcgccggg taaatacata taatacacaa
     1201 atctggctta ataaagtcta taatatatct cataaagaag tgctaaattg gctagtgcta
     1261 tatattttta agaaaatttc ttttgactaa gtccatatcg actttgtaaa agttcacatt
     1321 agcatacata tattacacga gccagaaata gtaacttttg cctaaatcac aaattgcaaa
     1381 atttaattgc ttgcaaaagg tcacatgctt ataatcaact tttttaaaaa tttaaaatac
     1441 ttttttattt tttattttta aacataaatg aaataattta tttattgttt atgattaccg
     1501 aaacataaaa cctgctcaag aaaaagaaac tgttttgtcc ttggaaaaaa agcactacct
     1561 aggagcggcc aaaatgccga ggctttcata gcttaaactc tttacagaaa ataggcatta
     1621 tagatcagtt cgagttttct tattcttcct tccggtttta tcgtcacagt tttacagtaa
     1681 ataagtatca cctcttagag ttcgatgata agctgtcaaa catgagaatt aattccacat
     1741 gttaaaatag tgaaggagca tgttcggcac acagtggacc gaacgtgggg taagtgcact
     1801 agggtccggt taaacggatc tcgcattgat gaggcaacgc taattatcaa catatagatt
     1861 gttatctatc tgcatgaaca cgaaatcttt acttgacgac ttgaggctga tggtgtttat
     1921 gcaaagaaac cactgtgttt aatatgtgtc actgtttgat attactgtca gcgtagaaga
     1981 taatagtaaa agcggttaat aagtgtattt gagataagtg tgataaagtt tttacagcga
     2041 aaagacgata aatacaagaa aatgattacg aggatacgga gagaggtatg tacatgtgta
     2101 tttatatact aagctgccgg cggttgtttg caagaccgag aaaaggctag caagaatcgg
     2161 gtcattgtag cgtatgcgcc tgtgaacatt ctcttcaaca agtttgattc cattgcggtg
     2221 aaatggtaaa agtcaacccc ctgcgatgta tattttcctg tacaatcaat caaaaagcca
     2281 aatgatttag cattatcttt acatcttgtt attttacaga ttttatgttt agatctttta
     2341 tgcttgcttt tcaaaaggct tgcaggcaag tgcacaaaca atacttaaat aaatactact
     2401 cagtaataac ctatttctta gcatttttga cgaaatttgc tattttgtta gagtctttta
     2461 caccatttgt ctccacacct ccgcttacat caacaccaat aacgccattt aatctaagcg
     2521 catcaccaac attttctggc gtcagtccac cagctaacat aaaatgtaag ctctcggggc
     2581 tctcttgcct tccaacccag tcagaaatcg agttccaatc caaaagttca cctgtcccac
     2641 ctgcttctga atcaaacaag ggaataaacg aatgaggttt ctgtgaagct gcactgagta
     2701 gtatgttgca gtcttttgga aatacgagtc ttttaataac tggcaaaccg aggaactctt
     2761 ggtattcttg ccacgactca tctccatgca gttggacgat cgatgataag ctgtcaaaca
     2821 tgagaattaa ttctacccta tgaacatatt ccattttgta atttcgtgtc gtttctatta
     2881 tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc tttttaagca
     2941 aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact gttggaacca
     3001 cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc ttcaatggcc
     3061 ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga caagatagtg
     3121 gcgatagggt tgaccttatt ctttggcaaa tctggagcag aaccgtggca tggttcgtac
     3181 aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg caacaaaccc
     3241 aaggaacctg ggataacgga ggcttcatcg gagatgatat caccaaacat gttgctggtg
     3301 attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc agaatcaatc
     3361 aattgatgtt gaaccttcaa tgtaggaaat tcgttcttga tggtttcctc cacagttttt
     3421 ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat aggcaatggt
     3481 ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac ttctggaacg
     3541 gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct cttaccaaag
     3601 taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc aaattgtggc
     3661 ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct taagttggcg
     3721 tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac actacctgta
     3781 ccccatttag gaccacccac agcacctaac aaaacggcat cagccttctt ggaggcttcc
     3841 agcgcctcat ctggaagtgg aacacctgta gcatcgatag cagcaccacc aattaaatga
     3901 ttttcgaaat cgaacttgac attggaacga acatcagaaa tagctttaag aaccttaatg
     3961 gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat cttcttaggg
     4021 gcagacatta gaatggtata tccttgaaat atatatatat attgctgaaa tgtaaaaggt
     4081 aagaaaagtt agaaagtaag acgattgcta accacctatt ggaaaaaaca ataggtcctt
     4141 aaataatatt gtcaacttca agtattgtga tgcaagcatt tagtcatgaa cgcttctcta
     4201 ttctatatga aaagccggtt ccggcgctct cacctttcct ttttctccca atttttcagt
     4261 tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca aaaaatttcc agtcatcgaa
     4321 tttgattctg tgcgatagcg cccctgtgtg ttctcgttat gttgaggaaa aaaataatgg
     4381 ttgctaagag attcgaactc ttgcatctta cgatacctga gtattcccac agttaattct
     4441 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4501 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4561 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4621 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4681 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4741 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4801 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4861 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     4921 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4981 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5041 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5101 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5161 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5221 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5281 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5341 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5401 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5461 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5521 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5581 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5641 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5701 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5761 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5821 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5881 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5941 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6001 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6061 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6121 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6181 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6241 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6301 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6361 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6421 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     6481 tggccgattc attaatgcag gatccgggat cgaagaaatg atggtaaatg aaataggaaa
     6541 tcaaggagca tgaaggcaaa agacaaatat aagggtcgaa cgaaaaataa agtgaaaagt
     6601 gttgatatga tgtatttggc tttgcggcgc cgaaaaaacg agtttacgca attgcacaat
     6661 catgctgact ctgtggcgga cccgcgctct tgccggcccg gcgataacgc tgggcgtgag
     6721 gctgtgcccg gcggagtttt ttgcgcctgc attttccaag gtttaccctg cgctaagggg
     6781 cgagattgga gaagcaataa gaatgccggt tggggttgcg atgatgacga ccacgacaac
     6841 tggtgtcatt atttaagttg ccgaaagaac ctgagtgcat ttgcaacatg agtatactag
     6901 aagaatgagc caagacttgc gagacgcgag tttgccggtg gtgcgaacaa tagagcgacc
     6961 atgaccttga aggtgagacg cgcataaccg ctagagtact ttgaagagga aacagcaata
     7021 gggttgctac cagtataaat agacaggtac atacaacact ggaaatggtt gtctgtttga
     7081 gtacgctttc aattcatttg ggtgtgcact ttattatgtt acaatatgga agggaacttt
     7141 acacttctcc tatgcacata tattaattaa agtccaatgc tagtagagaa ggggggtaac
     7201 acccctccgc gctcttttcc gatttttttc taaaccgtgg aatatttcgg atatcctttt
     7261 gttgtttccg ggtgtacaat atggacttcc tcttttctgg caaccaaacc catacatcgg
     7321 gattcctata ataccttcgt tggtctccct aacatgtagg tggcggaggg gagatataca
     7381 atagaacaga taccagacaa gacataatgg gctaaacaag actacaccaa ttacactgcc
     7441 tcattgatgg tggtacataa cgaactaata ctgtagccct agacttgata gccatcatca
     7501 tatcgaagtt tcactaccct ttttccattt gccatctatt gaagtaataa taggcgcatg
     7561 caacttcttt tctttttttt tcttttctct ctcccccgtt gttgtctcac catatccgca
     7621 atgacaaaaa aatgatggaa gacactaaag gaaaaaatta acgacaaaga cagcaccaac
     7681 agatgtcgtt gttccagagc tgatgagggg tatctcgaag cacacgaaac tttttccttc
     7741 cttcattcac gcacactact ctctaatgag caacggtata cggccttcct tccagttact
     7801 tgaatttgaa ataaaaaaaa gtttgctgtc ttgctatcaa gtataaatag acctgcaatt
     7861 attaatcttt tgtttcctcg tcattgttct cgttcccttt cttccttgtt tctttttctg
     7921 cacaatattt caagctatac caagcataca atcaactcca agctttgcaa agatggataa
     7981 agcggaatta attcccgagc ctccaaaaaa gaagagaaag gtcgaattgg gtaccgccgc
     8041 caattttaat caaagtggga atattgctga tagctcattg tccttcactt tcactaacag
     8101 tagcaacggt ccgaacctca taacaactca aacaaattct caagcgcttt cacaaccaat
     8161 tgcctcctct aacgttcatg ataacttcat gaataatgaa atcacggcta gtaaaattga
     8221 tgatggtaat aattcaaaac cactgtcacc tggttggacg gaccaaactg cgtataacgc
     8281 gtttggaatc actacaggga tgtttaatac cactacaatg gatgatgtat ataactatct
     8341 attcgatgat gaagataccc caccaaaccc aaaaaaagag atcgaattca tgacagacga
     8401 aactgctcac ccgacacaat ctgcttcaaa gcaggaatct gctgctctga aacagacagg
     8461 cgatgaccag caggaatctc agcaacaacg tggctacacc aactacaaca atggtagcaa
     8521 ctatacccaa aagaagcctt acaacagcaa cagacctcat cagcaaagag gtggtaaatt
     8581 tggaccaaac agatataaca accgtggcaa ctataatggt ggcggtagtt tcagaggtgg
     8641 acacatggga gccaacagct caaacgtgcc atggactggt tactataata actaccccgt
     8701 ttactaccag ccccagcaaa tggcggccgc tggtagtgcc cctgctaatc caattcctgt
     8761 cgaagagaag tcacctgttc caactaagat agaaatcact accaagtctg gtgaacactt
     8821 agatttgaaa gaacagcata aagccaagct acagtctcag gaaagatcta ctgtgtctcc
     8881 gcaaccagag tcaaagttaa aagaaacttc tgattctact tctacttcta ctccaactcc
     8941 taccccttcc actaatgact ctaaggccag ttctgaagaa aatatatctg aagctgaaaa
     9001 gacaagaaga aatttcatcg agcaagttaa acttcgtaaa gctgctttag aaaagaagag
     9061 aaaggagcaa cttgaaggtt ctagtggcaa caataatatt ccaatgaaga ctaccccaga
     9121 aaatgtggaa gagaagggat cggacaaacc tgaggtaacc gaaaaaacca agcctgctga
     9181 agaaaaatct gctgagccag aagttaagca agaaactcca gctgaagaag gtgaacaagg
     9241 agaaaagggc caaataaagg aagaatccac tccaaaggtg ttaacctttg ctgaacgttt
     9301 gaagttgaag aaacaacaaa aagaacgaga agaaaaaacg gaggggaagg aaaacaagga
     9361 agtgccagta caggaagaga ctaagagcgc catcgagtct gctcctgtgc cttga
//
