LOCUS       sequence-170817-        8136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Y2H Gal4 DNA binding domain for Cus2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8136)
  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 8136)
  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-CUS2
FEATURES             Location/Qualifiers
     source          1..8136
                     /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      2171..2218
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2245..2432
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2300..2324)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2456..2472)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2456..2472)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2469..2491)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2480..2496
                     /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(2504..2510,2511..2528,2529..2534))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2549..2570
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2687..2704)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2858..3446)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2938..2957)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3775..3822
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3847..3866
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4054..4848)
                     /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(4054..4848)
                     /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(4165..4184)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4775..4794
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4849..4953)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4980..6322
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5943..5990)
                     /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     6363..6381
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6419..6441)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6541..6560
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6581..6862
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6863..7537
                     /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(7640..8095)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7777..7798)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7989..8008
                     /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 catatggatg ctgatgaatt ggaattgaaa ggtcatttga
     1321 aaaaattgaa aaaagaagaa ttgcttagaa ggaaacagtt aaaagagagt aatcttcaga
     1381 aaagagagct tgaatataat aatgcctcaa aaaatacttc aatatatatt tctggtcttc
     1441 ccacagacaa aacaacgaaa gaaggactca ctgagcagtt ttgcaagtat ggaatgataa
     1501 gaaccaatcg agatggggaa cccctttgca agctatatgt gaatgataaa ggcgctttca
     1561 aaggggatgc cttgatcact tattccaagg aagaaagtgt tacattagca atcgaaatga
     1621 tgaacgagtc gatcttcctg ggaaaacaga tcagggtcga aagagcccaa tttcaaaaca
     1681 aggaaggtga caatatgcac gggaaagaaa atgatttaaa ggagttcaac ggacctgagc
     1741 ctccaataaa gaggctaaaa aaagcgaagt ctgaagggga gggggaagta atagattaca
     1801 atgacgatga aagcctagca aaagctgatc gaacggtgat atttgccaac gtattcaata
     1861 tttacaaatc atacacaaat gatgatataa atgatattca ggaagatctt ttggaaggct
     1921 gtgaagagat aggtcaagta gacagtatat ctgtatctcc caataagggg gaagcgacgg
     1981 tcgtctttaa gaacaataaa gttgcattgc aatgttgcaa aatcatgacc ggacgttatt
     2041 ttgatgggca aaagttgctt gctttcattt ctggagatga gaacacttca agcacaagtg
     2101 acaagaatga ggacagcgaa gtggaagatg accttatata gggatccgtc gacctgcagc
     2161 ggccgcataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgcg
     2221 cgcttgcagc caagctaatt ccgggcgaat ttcttatgat ttatgatttt tattattaaa
     2281 taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga
     2341 aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat
     2401 gaggtcgctc ttattgacca cacctctacc ggcatgcaag cttggcgtaa tcatggtcat
     2461 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2521 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2581 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2641 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2701 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2761 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2821 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2881 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2941 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3001 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3061 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3121 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3181 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3241 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3301 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3361 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3421 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3481 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3541 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3601 aacctgaggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct gggccttcac
     3661 ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg ccccaatggg
     3721 gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt ttatgaacaa
     3781 acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg cgggttcctt
     3841 ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg aagaactcca
     3901 gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc
     3961 ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg
     4021 cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata
     4081 gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc
     4141 ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg
     4201 gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat
     4261 gatattcggc aagcaggcat cgccatgagt cacgacgaga tcctcgccgt cgggcatgct
     4321 cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc
     4381 atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc
     4441 ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc
     4501 catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac
     4561 ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca
     4621 aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca gttcattcag
     4681 ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa
     4741 cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc
     4801 cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatac tcttcctttt
     4861 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4921 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4981 acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc
     5041 aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa gcgctatttt
     5101 accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat
     5161 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgagagcgct
     5221 attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat cccgagagcg
     5281 ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc tctataatgc
     5341 agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg ctactttggt
     5401 gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac tgattactag
     5461 cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc tataccgatg
     5521 tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc attggtcaga
     5581 aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa atgtttacat
     5641 tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt ttgtctaaag
     5701 agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca agttcaagga
     5761 gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata gcaaagagat
     5821 acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc gttacagtcc
     5881 ggtgcgtttt tggttttttg aaagtgcgtc ttcagagcgc ttttggtttt caaaagcgct
     5941 ctgaagttcc tatactttct agagaatagg aacttcggaa taggaacttc aaagcgtttc
     6001 cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca catacagctc actgttcacg
     6061 tcgcacctat atctgcgtgt tgcctgtata tatatataca tgagaagaac ggcatagtgc
     6121 gtgtttatgc ttaaatgcgt acttatatgc gtctatttat gtaggatgaa aggtagtcta
     6181 gtacctcctg tgatattatc ccattccatg cggggtatcg tatgcttcct tcagcactac
     6241 cctttagctg ttctatatgc tgccactcct caattggatt agtctcatcc ttcaatgcta
     6301 tcatttcctt tgatattgga tcatactaag aaaccattat tatcatgaca ttaacctata
     6361 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     6421 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     6481 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     6541 cggcatcaga gcagattgta ctgagagtgc accatagatc aacgacatta ctatatatat
     6601 aatataggaa gcatttaata gaacagcatc gtaatatatg tgtactttgc agttatgacg
     6661 ccagatggca gtagtggaag atattcttta ttgaaaaata gcttgtcacc ttacgtacaa
     6721 tcttgatccg gagcttttct ttttttgccg attaagaatt aattcggtcg aaaaaagaaa
     6781 aggagagggc caagagggag ggcattggtg actattgagc acgtgagtat acgtgattaa
     6841 gcacacaaag gcagcttgga gtatgtctgt tattaatttc acaggtagtt ctggtccatt
     6901 ggtgaaagtt tgcggcttgc agagcacaga ggccgcagaa tgtgctctag attccgatgc
     6961 tgacttgctg ggtattatat gtgtgcccaa tagaaagaga acaattgacc cggttattgc
     7021 aaggaaaatt tcaagtcttg taaaagcata taaaaatagt tcaggcactc cgaaatactt
     7081 ggttggcgtg tttcgtaatc aacctaagga ggatgttttg gctctggtca atgattacgg
     7141 cattgatatc gtccaactgc atggagatga gtcgtggcaa gaataccaag agttcctcgg
     7201 tttgccagtt attaaaagac tcgtatttcc aaaagactgc aacatactac tcagtgcagc
     7261 ttcacagaaa cctcattcgt ttattccctt gtttgattca gaagcaggtg ggacaggtga
     7321 acttttggat tggaactcga tttctgactg ggttggaagg caagagagcc ccgaaagctt
     7381 acattttatg ttagctggtg gactgacgcc agaaaatgtt ggtgatgcgc ttagattaaa
     7441 tggcgttatt ggtgttgatg taagcggagg tgtggagaca aatggtgtaa aagactctaa
     7501 caaaatagca aatttcgtca aaaatgctaa gaaataggtt attactgagt agtatttatt
     7561 taagtattgt ttgtgcactt gccgatctat gcggtgtgaa ataccgcaca gatgcgtaag
     7621 gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     7681 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     7741 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     7801 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     7861 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     7921 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     7981 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     8041 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt
     8101 cgccattcag gctgcgcaac tgttgggaag ggcgat
//
