LOCUS       sequence-183080-        7920 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Y2H Gal4 DNA binding domain for Hsh49.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7920)
  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 7920)
  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-HSH49
FEATURES             Location/Qualifiers
     source          1..7920
                     /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      1955..2002
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2029..2216
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2084..2108)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2240..2256)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2240..2256)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2253..2275)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2264..2280
                     /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(2288..2294,2295..2312,2313..2318))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2333..2354
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2471..2488)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2642..3230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2722..2741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3559..3606
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3631..3650
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3838..4632)
                     /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(3838..4632)
                     /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(3949..3968)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4559..4578
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4633..4737)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4764..6106
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5727..5774)
                     /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     6147..6165
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6203..6225)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6325..6344
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6365..6646
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6647..7321
                     /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(7424..7879)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7561..7582)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7773..7792
                     /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 catatgaact acagcgcaga ctctggtaat acagtttatg
     1321 tggggaatat agatccaagg ataacgaaag aacaacttta tgaactcttc attcagatca
     1381 accctgtttt aaggattaag tatccgaaag ataaagttct acaagcttac caaggctacg
     1441 cattcatcga attttataat caaggagatg ctcaatatgc aataaaaata atgaataata
     1501 ccgttagatt atacgaccga ctgataaagg ttcgtcaagt aaccaattct acaggaacaa
     1561 ctaatttacc ttcaaatatt tctaaggaca tgattctacc tattgctaag ttgttcataa
     1621 aaaaccttgc agattcgata gatagcgacc aattagtgaa aattttcaat aaatttggta
     1681 agctcataag ggaacccgaa attttctacc tatcaaacgg aaaattaaag tgtgcttatg
     1741 tttattttga ggactttgaa aaagcagatt tggcaataaa atctttgaac aatcagttag
     1801 tcgccaacaa cagaattact gttgattatg ccttcaaaga aaatggaaaa ggtaacgcaa
     1861 aatacggtga tgacgttgac aggctgttga ataaagaagc attaaagcac aacatgttga
     1921 aataaggatc cgtcgacctg cagcggccgc ataactagca taaccccttg gggcctctaa
     1981 acgggtcttg aggggttttt tgcgcgcttg cagccaagct aattccgggc gaatttctta
     2041 tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa
     2101 agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc
     2161 aggttgcttt ctcaggtata gcatgaggtc gctcttattg accacacctc taccggcatg
     2221 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2281 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2341 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2401 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2461 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2521 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2581 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2641 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2701 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2761 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2821 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2881 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2941 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3001 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3061 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3121 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3181 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3241 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3301 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3361 aatcaatcta aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt
     3421 tggctgcgag ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa
     3481 cgcgggcgta ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg
     3541 accgaacccc gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta
     3601 ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc
     3661 ccctagggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg
     3721 cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat
     3781 ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca
     3841 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
     3901 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
     3961 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     4021 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gagtcacgac
     4081 gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag
     4141 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     4201 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     4261 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
     4321 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     4381 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     4441 tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     4501 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     4561 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     4621 ttgttcaatc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4681 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4741 atttccccga aaagtgccac ctgaacgaag catctgtgct tcattttgta gaacaaaaat
     4801 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     4861 aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca
     4921 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     4981 acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt
     5041 ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt
     5101 tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta
     5161 aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca
     5221 cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat
     5281 ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc
     5341 gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat
     5401 actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc
     5461 ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt
     5521 cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag
     5581 cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa
     5641 tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcttcaga
     5701 gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc
     5761 ggaataggaa cttcaaagcg tttccgaaaa cgagcgcttc cgaaaatgca acgcgagctg
     5821 cgcacataca gctcactgtt cacgtcgcac ctatatctgc gtgttgcctg tatatatata
     5881 tacatgagaa gaacggcata gtgcgtgttt atgcttaaat gcgtacttat atgcgtctat
     5941 ttatgtagga tgaaaggtag tctagtacct cctgtgatat tatcccattc catgcggggt
     6001 atcgtatgct tccttcagca ctacccttta gctgttctat atgctgccac tcctcaattg
     6061 gattagtctc atccttcaat gctatcattt cctttgatat tggatcatac taagaaacca
     6121 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     6181 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     6241 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     6301 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     6361 gatcaacgac attactatat atataatata ggaagcattt aatagaacag catcgtaata
     6421 tatgtgtact ttgcagttat gacgccagat ggcagtagtg gaagatattc tttattgaaa
     6481 aatagcttgt caccttacgt acaatcttga tccggagctt ttcttttttt gccgattaag
     6541 aattaattcg gtcgaaaaaa gaaaaggaga gggccaagag ggagggcatt ggtgactatt
     6601 gagcacgtga gtatacgtga ttaagcacac aaaggcagct tggagtatgt ctgttattaa
     6661 tttcacaggt agttctggtc cattggtgaa agtttgcggc ttgcagagca cagaggccgc
     6721 agaatgtgct ctagattccg atgctgactt gctgggtatt atatgtgtgc ccaatagaaa
     6781 gagaacaatt gacccggtta ttgcaaggaa aatttcaagt cttgtaaaag catataaaaa
     6841 tagttcaggc actccgaaat acttggttgg cgtgtttcgt aatcaaccta aggaggatgt
     6901 tttggctctg gtcaatgatt acggcattga tatcgtccaa ctgcatggag atgagtcgtg
     6961 gcaagaatac caagagttcc tcggtttgcc agttattaaa agactcgtat ttccaaaaga
     7021 ctgcaacata ctactcagtg cagcttcaca gaaacctcat tcgtttattc ccttgtttga
     7081 ttcagaagca ggtgggacag gtgaactttt ggattggaac tcgatttctg actgggttgg
     7141 aaggcaagag agccccgaaa gcttacattt tatgttagct ggtggactga cgccagaaaa
     7201 tgttggtgat gcgcttagat taaatggcgt tattggtgtt gatgtaagcg gaggtgtgga
     7261 gacaaatggt gtaaaagact ctaacaaaat agcaaatttc gtcaaaaatg ctaagaaata
     7321 ggttattact gagtagtatt tatttaagta ttgtttgtgc acttgccgat ctatgcggtg
     7381 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat
     7441 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     7501 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     7561 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     7621 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     7681 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     7741 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     7801 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     7861 gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg gaagggcgat
//
