LOCUS       sequence-170810-        8079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Y2H Gal4 DNA binding domain for Bud13.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8079)
  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 8079)
  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-BUD13
FEATURES             Location/Qualifiers
     source          1..8079
                     /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      2114..2161
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2188..2375
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2243..2267)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2399..2415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2399..2415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2412..2434)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2423..2439
                     /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(2447..2453,2454..2471,2472..2477))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2492..2513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2630..2647)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2801..3389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2881..2900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3718..3765
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3790..3809
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3997..4791)
                     /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(3997..4791)
                     /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(4108..4127)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4718..4737
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4792..4896)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4923..6265
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5886..5933)
                     /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     6306..6324
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6362..6384)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6484..6503
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6524..6805
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6806..7480
                     /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(7583..8038)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7720..7741)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7932..7951
                     /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 catatggcat tgcatcagta tttatcagag acttatgggc
     1321 ccacgaaacc caaaaataag acgaaaaaga agaagaaaga gtcaaaatca gacgctaact
     1381 cagacaaaac ttctttgata gtaaaagaac ggctaagtac actgcaacaa gaacaggaga
     1441 agtcaggagt tgcttcattc agcaagtttg acaaacaaaa aagcaagaat atatggaaga
     1501 acctggaaac aaacgagctt tcccatgcaa taacacatcc ttccgcatcg tcaattactg
     1561 gcaacgaaag caagaacgat ctaaaggaaa tcagggctca agagccactt gtcacagtag
     1621 cagacaaatc gaaaacacga aaaaccatat acagagacgc tcaaggtcac aagattcagg
     1681 aagattccaa gatagacgat tctagtttta gtcgatctaa atatgaagat gagaaagccg
     1741 cggaaagaga gcaatacctg aaaaatttga atatgggaga cgtgcaaaag cttggaataa
     1801 atgtagatgc acatgataag aagaaaaatc aaactgcctc gagtctgacg atagaagacc
     1861 ctgcaataac atttacacat gacaaagaaa gaactgtaaa aacatcttta ctgggccgca
     1921 agctttatga taagccagca cctgagaaca ggtttgccat tatgcctggg tcaagatggg
     1981 acggtgtcca cagatcaaat ggctttgaag aaaaatggtt tgctaagcaa aatgagatca
     2041 atgagaagaa agtgcaaagc tacaccctac aggaggatta ttgaggatcc gtcgacctgc
     2101 agcggccgca taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     2161 gcgcgcttgc agccaagcta attccgggcg aatttcttat gatttatgat ttttattatt
     2221 aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa
     2281 cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag
     2341 catgaggtcg ctcttattga ccacacctct accggcatgc aagcttggcg taatcatggt
     2401 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     2461 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     2521 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     2581 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     2641 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2701 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2761 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2821 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2881 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2941 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3001 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3061 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3121 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3181 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3241 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3301 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3361 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3421 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3481 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3541 agtaacctga ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt
     3601 cacccgaact tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat
     3661 ggggtctcgg tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa
     3721 caaacgaccc aacaccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc
     3781 cttccggtat tgtctccttc cgtgtttcag ttagcctccc cctagggtgg gcgaagaact
     3841 ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga
     3901 agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg
     3961 tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     4021 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     4081 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     4141 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     4201 catgatattc ggcaagcagg catcgccatg agtcacgacg agatcctcgc cgtcgggcat
     4261 gctcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     4321 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     4381 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     4441 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     4501 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     4561 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt
     4621 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     4681 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     4741 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tactcttcct
     4801 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4861 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4921 tgaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt
     4981 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat
     5041 tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct
     5101 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc
     5161 gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga
     5221 gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa
     5281 tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt
     5341 ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac
     5401 tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg
     5461 atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc
     5521 agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta
     5581 cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta
     5641 aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa
     5701 ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga
     5761 gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag
     5821 tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag cgcttttggt tttcaaaagc
     5881 gctctgaagt tcctatactt tctagagaat aggaacttcg gaataggaac ttcaaagcgt
     5941 ttccgaaaac gagcgcttcc gaaaatgcaa cgcgagctgc gcacatacag ctcactgttc
     6001 acgtcgcacc tatatctgcg tgttgcctgt atatatatat acatgagaag aacggcatag
     6061 tgcgtgttta tgcttaaatg cgtacttata tgcgtctatt tatgtaggat gaaaggtagt
     6121 ctagtacctc ctgtgatatt atcccattcc atgcggggta tcgtatgctt ccttcagcac
     6181 taccctttag ctgttctata tgctgccact cctcaattgg attagtctca tccttcaatg
     6241 ctatcatttc ctttgatatt ggatcatact aagaaaccat tattatcatg acattaacct
     6301 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     6361 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     6421 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     6481 atgcggcatc agagcagatt gtactgagag tgcaccatag atcaacgaca ttactatata
     6541 tataatatag gaagcattta atagaacagc atcgtaatat atgtgtactt tgcagttatg
     6601 acgccagatg gcagtagtgg aagatattct ttattgaaaa atagcttgtc accttacgta
     6661 caatcttgat ccggagcttt tctttttttg ccgattaaga attaattcgg tcgaaaaaag
     6721 aaaaggagag ggccaagagg gagggcattg gtgactattg agcacgtgag tatacgtgat
     6781 taagcacaca aaggcagctt ggagtatgtc tgttattaat ttcacaggta gttctggtcc
     6841 attggtgaaa gtttgcggct tgcagagcac agaggccgca gaatgtgctc tagattccga
     6901 tgctgacttg ctgggtatta tatgtgtgcc caatagaaag agaacaattg acccggttat
     6961 tgcaaggaaa atttcaagtc ttgtaaaagc atataaaaat agttcaggca ctccgaaata
     7021 cttggttggc gtgtttcgta atcaacctaa ggaggatgtt ttggctctgg tcaatgatta
     7081 cggcattgat atcgtccaac tgcatggaga tgagtcgtgg caagaatacc aagagttcct
     7141 cggtttgcca gttattaaaa gactcgtatt tccaaaagac tgcaacatac tactcagtgc
     7201 agcttcacag aaacctcatt cgtttattcc cttgtttgat tcagaagcag gtgggacagg
     7261 tgaacttttg gattggaact cgatttctga ctgggttgga aggcaagaga gccccgaaag
     7321 cttacatttt atgttagctg gtggactgac gccagaaaat gttggtgatg cgcttagatt
     7381 aaatggcgtt attggtgttg atgtaagcgg aggtgtggag acaaatggtg taaaagactc
     7441 taacaaaata gcaaatttcg tcaaaaatgc taagaaatag gttattactg agtagtattt
     7501 atttaagtat tgtttgtgca cttgccgatc tatgcggtgt gaaataccgc acagatgcgt
     7561 aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta
     7621 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     7681 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     7741 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     7801 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     7861 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     7921 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     7981 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     8041 attcgccatt caggctgcgc aactgttggg aagggcgat
//
