LOCUS       sequence-170833-        8427 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Y2H Gal4 DNA binding domain for Slu7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8427)
  AUTHORS   Carrocci TJ, Zoerner DM, Paulson JC, Hoskins AA
  TITLE     SF3b1 mutations associated with myelodysplastic syndromes alter the 
            fidelity of branchsite selection in yeast.
  JOURNAL   Nucleic Acids Res. 2017 Jan 6. pii: gkw1349. doi: 
            10.1093/nar/gkw1349.
  PUBMED    28062854
REFERENCE   2  (bases 1 to 8427)
  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: "Nucleic 
            Acids Res. 2017 Jan 6. pii: gkw1349. doi: 10.1093/nar/gkw1349."
COMMENT     Sequence Label: pGBKT7-SLU7
FEATURES             Location/Qualifiers
     source          1..8427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..734
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             762..1202
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     primer_bind     1213..1232
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1213..1231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1251..1280
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      2462..2509
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2536..2723
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2591..2615)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2747..2763)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2747..2763)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2760..2782)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2771..2787
                     /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)."
     promoter        complement(join(2795..2801,2802..2819,2820..2825))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2840..2861
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2978..2995)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3149..3737)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3229..3248)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4066..4113
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4138..4157
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4345..5139)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4345..5139)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4456..4475)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5066..5085
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5140..5244)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5271..6613
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(6234..6281)
                     /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)."
     primer_bind     6654..6672
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6710..6732)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6832..6851
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6872..7153
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             7154..7828
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(7931..8386)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8068..8089)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8280..8299
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cggtgcgggc ctcttcgcta ttacgccaga tccttttgtt gtttccgggt gtacaatatg
       61 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
      121 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
      181 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
      241 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
      301 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
      361 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
      421 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
      481 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
      541 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
      601 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
      661 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
      721 gcatacaatc aactccaagc ttgaagcaag cctcctgaaa gatgaagcta ctgtcttcta
      781 tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga
      841 agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt
      901 ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc
      961 tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac
     1021 aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgccg
     1081 tcacagatag attggcttca gtggagactg atatgcctct aacattgaga cagcatagaa
     1141 taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag ttgactgtat
     1201 cgccggaatt tgtaatacga ctcactatag ggcgagccgc catcatggag gagcagaagc
     1261 tgatctcaga ggaggacctg catatgaata ataacagcag aaacaacgaa aatcgaagca
     1321 ctattaacag aaataaaagg caactacaac aagcaaaaga aaaaaatgaa aatattcata
     1381 tccccaggta tattagaaat caaccatggt actataagga tacccccaaa gaacaagaag
     1441 ggaagaagcc cggcaatgat gatacgagca ctgcagaagg aggagaaaaa agcgactact
     1501 tggtgcatca taggcaaaaa gcaaaagggg gtgctttaga tattgacaat aattcagaac
     1561 caaaaattgg tatgggtata aaggatgagt tcaaactaat cagaccccag aagatgtccg
     1621 tccgagattc tcattcgctg tcattttgta ggaattgtgg ggaagcaggg cataaggaga
     1681 aagactgcat ggaaaaacct cgtaagatgc agaagcttgt tcccgattta aattcacaaa
     1741 aaaataatgg cacagtttta gtacgagcta ctgatgatga ctgggactcc agaaaagata
     1801 gatggtacgg ttactcaggg aaagaataca atgaactgat aagtaagtgg gagcgtgata
     1861 aaagaaataa aataaaagga aaagacaaat cccaaactga tgaaacacta tgggatacag
     1921 atgaagagat agaactaatg aagttagaac tttacaagga ttccgtaggt tcattgaaga
     1981 aagatgatgc tgataattct cagttgtata ggacatcaac gagattgaga gaagataagg
     2041 ctgcttactt gaacgacata aattcaacgg agagtaatta tgatcctaaa tcaagattgt
     2101 acaaaactga aacactgggc gcagttgatg aaaaatcaaa aatgttccgc agacatttga
     2161 caggtgaagg cctaaaatta aacgaattga accagtttgc tagatctcac gctaaggaaa
     2221 tgggtatacg tgatgaaatt gaggataagg aaaaagtaca acatgtttta gtcgccaatc
     2281 ctactaaata tgaatatctg aagaaaaaac gggaacaaga agaaaccaag cagcccaaga
     2341 ttgtcagcat tggagatctg gaagctagga aagtagatgg tacaaagcaa tctgaggaac
     2401 aacggaacca cttaaaagat ttatatggtt aaggatccgt cgacctgcag cggccgcata
     2461 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc gcgcttgcag
     2521 ccaagctaat tccgggcgaa tttcttatga tttatgattt ttattattaa ataagttata
     2581 aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta
     2641 ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagca tgaggtcgct
     2701 cttattgacc acacctctac cggcatgcaa gcttggcgta atcatggtca tagctgtttc
     2761 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     2821 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2881 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2941 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3001 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3061 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3121 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3181 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3241 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3301 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3361 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3421 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3481 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3541 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3601 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3661 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3721 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3781 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3841 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taacctgagg
     3901 ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc tgggccttca cccgaacttg
     3961 gggggtgggg tggggaaaag gaagaaacgc gggcgtattg gccccaatgg ggtctcggtg
     4021 gggtatcgac agagtgccag ccctgggacc gaaccccgcg tttatgaaca aacgacccaa
     4081 caccgtgcgt tttattctgt ctttttattg ccgtcatagc gcgggttcct tccggtattg
     4141 tctccttccg tgtttcagtt agcctccccc tagggtgggc gaagaactcc agcatgagat
     4201 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
     4261 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
     4321 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
     4381 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
     4441 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
     4501 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
     4561 caagcaggca tcgccatgag tcacgacgag atcctcgccg tcgggcatgc tcgccttgag
     4621 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
     4681 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
     4741 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     4801 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     4861 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     4921 cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc agttcattca gggcaccgga
     4981 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     5041 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     5101 ggccggagaa cctgcgtgca atccatcttg ttcaatcata ctcttccttt ttcaatatta
     5161 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5221 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aacgaagcat
     5281 ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     5341 atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa
     5401 gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     5461 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca
     5521 acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc
     5581 taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg
     5641 ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt
     5701 ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc
     5761 gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg
     5821 catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga
     5881 acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt
     5941 gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact
     6001 agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt
     6061 ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag
     6121 caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt
     6181 ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc tctgaagttc
     6241 ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt ccgaaaacga
     6301 gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac gtcgcaccta
     6361 tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg cgtgtttatg
     6421 cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct agtacctcct
     6481 gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta ccctttagct
     6541 gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct atcatttcct
     6601 ttgatattgg atcatactaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     6661 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     6721 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     6781 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     6841 agcagattgt actgagagtg caccatagat caacgacatt actatatata taatatagga
     6901 agcatttaat agaacagcat cgtaatatat gtgtactttg cagttatgac gccagatggc
     6961 agtagtggaa gatattcttt attgaaaaat agcttgtcac cttacgtaca atcttgatcc
     7021 ggagcttttc tttttttgcc gattaagaat taattcggtc gaaaaaagaa aaggagaggg
     7081 ccaagaggga gggcattggt gactattgag cacgtgagta tacgtgatta agcacacaaa
     7141 ggcagcttgg agtatgtctg ttattaattt cacaggtagt tctggtccat tggtgaaagt
     7201 ttgcggcttg cagagcacag aggccgcaga atgtgctcta gattccgatg ctgacttgct
     7261 gggtattata tgtgtgccca atagaaagag aacaattgac ccggttattg caaggaaaat
     7321 ttcaagtctt gtaaaagcat ataaaaatag ttcaggcact ccgaaatact tggttggcgt
     7381 gtttcgtaat caacctaagg aggatgtttt ggctctggtc aatgattacg gcattgatat
     7441 cgtccaactg catggagatg agtcgtggca agaataccaa gagttcctcg gtttgccagt
     7501 tattaaaaga ctcgtatttc caaaagactg caacatacta ctcagtgcag cttcacagaa
     7561 acctcattcg tttattccct tgtttgattc agaagcaggt gggacaggtg aacttttgga
     7621 ttggaactcg atttctgact gggttggaag gcaagagagc cccgaaagct tacattttat
     7681 gttagctggt ggactgacgc cagaaaatgt tggtgatgcg cttagattaa atggcgttat
     7741 tggtgttgat gtaagcggag gtgtggagac aaatggtgta aaagactcta acaaaatagc
     7801 aaatttcgtc aaaaatgcta agaaataggt tattactgag tagtatttat ttaagtattg
     7861 tttgtgcact tgccgatcta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     7921 ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     7981 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     8041 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     8101 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     8161 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     8221 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     8281 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     8341 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca
     8401 ggctgcgcaa ctgttgggaa gggcgat
//
