LOCUS       sequence-182740-        8392 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express yeast Kre33 with Y2H binding domain fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8392)
  AUTHORS   Vincent NG, Charette JM, Baserga SJ
  TITLE     The SSU processome interactome in Saccharomyces cerevisiae reveals 
            potential new protein subcomplexes.
  JOURNAL   RNA. 2017 Oct 20. pii: rna.062927.117. doi: 10.1261/rna.062927.117.
  PUBMED    29054886
REFERENCE   2  (bases 1 to 8392)
  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: "RNA. 2017 
            Oct 20. pii: rna.062927.117. doi: 10.1261/rna.062927.117."
COMMENT     Sequence Label: pAS2-1-Kre33
FEATURES             Location/Qualifiers
     source          1..8392
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     rep_origin      2..1344
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(965..1012)
                     /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     1385..1403
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1441..1463)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1563..1582
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1603..1883
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             1884..2558
                     /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(2661..3116)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2798..2819)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3010..3029
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3246..3268
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3260..3277
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3261..3277
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(3314..3561)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3527..3545
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        4769..5473
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             5501..5941
                     /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"
     terminator      6030..6217
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6085..6109)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(6241..6257)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6241..6257)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6254..6276)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6265..6281
                     /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(6289..6295,6296..6313,6314..6319))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(6472..6489)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6643..7231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6723..6742)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7402..8193,8194..8262))
                     /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     8025..8044
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8263..8367)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
       61 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
      121 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
      181 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
      241 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
      301 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
      361 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
      421 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
      481 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
      541 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
      601 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
      661 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
      721 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
      781 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
      841 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
      901 ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg
      961 ctctgaagtt cctatacttt ctagagaata ggaacttcgg aataggaact tcaaagcgtt
     1021 tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg cacatacagc tcactgttca
     1081 cgtcgcacct atatctgcgt gttgcctgta tatatatata catgagaaga acggcatagt
     1141 gcgtgtttat gcttaaatgc gtacttatat gcgtctattt atgtaggatg aaaggtagtc
     1201 tagtacctcc tgtgatatta tcccattcca tgcggggtat cgtatgcttc cttcagcact
     1261 accctttagc tgttctatat gctgccactc ctcaattgga ttagtctcat ccttcaatgc
     1321 tatcatttcc tttgatattg gatcatatta agaaaccatt attatcatga cattaaccta
     1381 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     1441 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     1501 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     1561 tgcggcatca gagcagattg tactgagagt gcaccataga tcaacgacat tactatatat
     1621 ataatatagg aagcatttaa tagacagcat cgtaatatat gtgtactttg cagttatgac
     1681 gccagatggc agtagtggaa gatattcttt attgaaaaat agcttgtcac cttacgtaca
     1741 atcttgatcc ggagcttttc tttttttgcc gattaagaat taattcggtc gaaaaaagaa
     1801 aaggagaggg ccaagaggga gggcattggt gactattgag cacgtgagta tacgtgatta
     1861 agcacacaaa ggcagcttgg agtatgtctg ttattaattt cacaggtagt tctggtccat
     1921 tggtgaaagt ttgcggcttg cagagcacag aggccgcaga atgtgctcta gattccgatg
     1981 ctgacttgct gggtattata tgtgtgccca atagaaagag aacaattgac ccggttattg
     2041 caaggaaaat ttcaagtctt gtaaaagcat ataaaaatag ttcaggcact ccgaaatact
     2101 tggttggcgt gtttcgtaat caacctaagg aggatgtttt ggctctggtc aatgattacg
     2161 gcattgatat cgtccaactg catggagatg agtcgtggca agaataccaa gagttcctcg
     2221 gtttgccagt tattaaaaga ctcgtatttc caaaagactg caacatacta ctcagtgcag
     2281 cttcacagaa acctcattcg tttattccct tgtttgattc agaagcaggt gggacaggtg
     2341 aacttttgga ttggaactcg atttctgact gggttggaag gcaagagagc cccgaaagct
     2401 tacattttat gttagctggt ggactgacgc cagaaaatgt tggtgatgcg cttagattaa
     2461 atggcgttat tggtgttgat gtaagcggag gtgtggagac aaatggtgta aaagactcta
     2521 acaaaatagc aaatttcgtc aaaaatgcta agaaataggt tattactgag tagtatttat
     2581 ttaagtattg tttgtgcact tgccgatcta tgcggtgtga aataccgcac agatgcgtaa
     2641 ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
     2701 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2761 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2821 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2881 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2941 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3001 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3061 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcgcg
     3121 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     3181 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     3241 gttttcccag tcacgacgtt gtaaaacgac ggccagtcgt ccaagctttc gcgagctcga
     3301 gatcccgagc tttgcaaatt aaagccttcg agcgtcccaa aaccttctca agcaaggttt
     3361 tcagtataat gttacatgcg tacacgcgtc tgtacagaaa aaaaagaaaa atttgaaata
     3421 taaataacgt tcttaatact aacataacta taaaaaaata aatagggacc tagacttcag
     3481 gttgtctaac tccttccttt tcggttagag cggatgtggg gggagggcgt gaatgtaagc
     3541 gtgacataac taattacatg atatcgacaa aggaaaaggg gcctgtttac tcacaggctt
     3601 ttttcaagta ggtaattaag tcgtttctgt ctttttcctt cttcaaccca ccaaaggcca
     3661 tcttggtact tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt
     3721 tttttttttt tttttttttt tttttttttt tttttttttt tttcatagaa ataatacaga
     3781 agtagatgtt gaattagatt aaactgaaga tatataattt attggaaaat acatagagct
     3841 ttttgttgat gcgcttaagc gatcaattca acaacaccac cagcagctct gattttttct
     3901 tcagccaact tggagacgaa tctagctttg acgataactg gaacatttgg gattctaccc
     3961 ttacccaaga tcttaccgta accggctgcc aaagtgtcaa taactggagc agtttcctta
     4021 gaagcagatt tcaagtattg gtctctcttg tcttctggga tcaatgtcca caatttgtcc
     4081 aagttcaaga ctggcttcca gaaatgagct tgttgcttgt ggaagtatct cataccaacc
     4141 ttaccgaaat aacctggatg gtatttatcc atgttaattc tgtggtgatg ttgaccaccg
     4201 gccatacctc taccaccggg gtgctttctg tgcttaccga tacgaccttt accggctgag
     4261 acgtgacctc tgtgctttct agtcttagtg aatctggaag gcattcttga ttagttggat
     4321 gattgttctg ggatttaatg caaaaaaatc actaagaagg aaaaaaatca acggagaaag
     4381 caaacgccat cttaaatata cgggatacag atgaaaggtt tgaacctatc tgggaaaata
     4441 cgcattaaac aagcgaaaaa ctgcgaggaa aattgtttgc gtctctgcgg gctattcacg
     4501 cgccagagga aaataggaaa aataacaggg cattagaaaa ataattttga ttttggtaat
     4561 gtgtgggtcc ctggtgtaca gatgttacat tggttacagt actcttgttt ttgctgtgtt
     4621 tttcgatgaa tctccaaaat ggttgttagc acatggaaga gtcaccgatg ctaagttatc
     4681 tctatgtaag ctacgtggcg tgacttttga tgaagccgca caagagatac aggattggca
     4741 actgcaaata gaatctgggg atctagatat ccttttgttg tttccgggtg tacaatatgg
     4801 acttcctctt ttctggcaac caaacccata catcgggatt cctataatac cttcgttggt
     4861 ctccctaaca tgtaggtggc ggaggggaga tatacaatag aacagatacc agacaagaca
     4921 taatgggcta aacaagacta caccaattac actgcctcat tgatggtggt acataacgaa
     4981 ctaatactgt agccctagac ttgatagcca tcatcatatc gaagtttcac tacccttttt
     5041 ccatttgcca tctattgaag taataatagg cgcatgcaac ttcttttctt tttttttctt
     5101 ttctctctcc cccgttgttg tctcaccata tccgcaatga caaaaaaaat gatggaagac
     5161 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
     5221 tgaggggtat cttcgaacac acgaaacttt ttccttcctt cattcacgca cactactctc
     5281 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaagttt
     5341 gccgctttgc tatcaagtat aaatagacct gcaattatta atcttttgtt tcctcgtcat
     5401 tgttctcgtt ccctttcttc cttgtttctt tttctgcaca atatttcaag ctataccaag
     5461 catacaatca actccaagct tgaagcaagc ctcctgaaag atgaagctac tgtcttctat
     5521 cgaacaagca tgcgatattt gccgacttaa aaagctcaag tgctccaaag aaaaaccgaa
     5581 gtgcgccaag tgtctgaaga acaactggga gtgtcgctac tctcccaaaa ccaaaaggtc
     5641 tccgctgact agggcacatc tgacagaagt ggaatcaagg ctagaaagac tggaacagct
     5701 atttctactg atttttcctc gagaagacct tgacatgatt ttgaaaatgg attctttaca
     5761 ggatataaaa gcattgttaa caggattatt tgtacaagat aatgtgaata aagatgccgt
     5821 cacagataga ttggcttcag tggagactga tatgcctcta acattgagac agcatagaat
     5881 aagtgcgaca tcatcatcgg aagagagtag taacaaaggt caaagacagt tgactgtatc
     5941 gccggtattg caatacccag ctttgactca tatggccatg gaggccgaat tcccggggat
     6001 ccgtcgacct gcagccaagc taattccggg cgaatttctt atgatttatg atttttatta
     6061 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
     6121 aacgaaaatt cttgttcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
     6181 agcatgaggt cgctcttatt gaccacacct ctaccggcat gcaagcttgg cgtaatcatg
     6241 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6301 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg aggtaactca cattaattgc
     6361 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgg attaatgaat
     6421 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6481 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6541 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6601 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6661 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6721 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6781 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6841 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6901 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6961 cccggtaaga cacgacttat cgccactggc agcagccact agtaacagga ttagcagagc
     7021 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7081 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7141 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7201 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7261 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7321 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7381 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7441 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7501 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7561 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7621 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7681 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7741 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7801 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7861 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7921 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7981 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8041 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8101 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8161 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8221 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8281 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8341 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ct
//
