LOCUS       sequence-104523-        9411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integration plasmid targeted to the HO loci with heterodimeric
            Hsp82. Digest with NotI prior to addition to yeast..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9411)
  AUTHORS   Mishra P, Bolon DN
  TITLE     Designed Hsp90 heterodimers reveal an asymmetric ATPase-driven 
            mechanism in vivo.
  JOURNAL   Mol Cell. 2014 Jan 23;53(2):344-50. doi: 
            10.1016/j.molcel.2013.12.024.
  PUBMED    24462207
REFERENCE   2  (bases 1 to 9411)
  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"; doi: 
            "10.1016/j.molcel.2013.12"; journalName: "Mol Cell"; date: 
            "2014-01-23- 23"; volume: "53"; issue: "2"; pages: "344-50"
COMMENT     Sequence Label: pHOHO2_Hsp82viiA
FEATURES             Location/Qualifiers
     source          1..9411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        962..1359
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1384..1401
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3700..3716)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3701..3717)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      3719..3966
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3735..3753)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(3985..4003)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4013..4029)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4174..4629
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(4889..5548)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(5549..5736)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     promoter        complement(6433..6451)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6458..6474)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7081..7100)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7200..7222
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7260..7278)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7346..7450
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7451..7519,7520..8311)
                     /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     complement(7669..7688)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8482..9070
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8971..8990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9224..9241
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9358..9379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tcgagatccg atatcgccgt ggcggccgcc agctgaagct taattatcct gggcacgagt
       61 gaaacaaagc taaaaccttt atttagcatg gccattgaat gtaacaatta tatatatcgc
      121 aagcacaaaa aatcaaggag agagaactac cactttgttc atgtgtacaa tgttcattat
      181 ctccataagc aaaaaaaaaa aaatagaaaa catatgctat aaggttgata ttctcacgag
      241 taagcggcac ttgctactta ttgacattgc agatttttgg ctacagaaat agtatattag
      301 agattataat tgctaatcaa atcaaaatat aaaattagta aaccaaacca tttataccct
      361 tccttagtag ttatggattg ttttttaatg atatttctgc aaaccaaaga aagattgtta
      421 tccagataga atttagtttt gatattcatt tttttgttga agattgaacg ccatatctgg
      481 gcctcataat tcaaaagacg gtgccattat cggtagcgtt tcgcattgta ctggatttca
      541 gaaatttcac agttgatgaa tcgaaaagaa tggtctcatt gcaacacgta aggttaagat
      601 gtcccttttt accattatag gcaataaatg aatcataaaa cgaccgtata ctggtgaaat
      661 agtagggaga acgagtacct gtagtaaaaa gtataaatca tagttaatcg ggcaatgtcc
      721 ctcgatcaag gagtattgtg tcatgttcga gacaaacgcc aacatttttg tttcttttgg
      781 acaaatgttg tttgcattta tgatccgtta tattttgatc taatgtagag ttgcacgtag
      841 ttcttactgg caaagaaatc gatgcatacc aaaaaagaat aaaggtgata tttgatcttt
      901 accgtttagt tccaacgtaa aattgtgcct ttggacttaa aatggcgtga gctcagttta
      961 tatagcttca aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc
     1021 atcgccgtac cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct
     1081 ctagggtgtc gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt
     1141 ttctttttct tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa
     1201 tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg
     1261 gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt
     1321 cttgctcatt agaaagaaag catagcaatc taatctaagt tttctagaac tagtatgggt
     1381 ggtcaccacc atcaccacca cggtggtcat atggctagtg aaacttttga atttcaagct
     1441 gaaattactc agttgatgag tttgatcatc aacaccgtct attctaacaa ggaaattttc
     1501 ttgagagaac tgatatctaa tgcctcggat gcgttggata aaattagata caaatctttg
     1561 tctgatccaa agcaattgga aacagaacca gatctcttta ttagaatcac tccaaagcca
     1621 gagcaaaaag ttttggaaat cagagattct ggtattggta tgaccaaggc tgaattgatt
     1681 aataacttgg gtaccattgc caagtctggt accaaagcct tcatggaagc tctatctgct
     1741 ggtgccgatg tatccatgat tggtcaattc ggtgttggtt tttactcttt attcttagtt
     1801 gccgacagag ttcaggttat ttcaaagagc aacgacgacg aacaatacat ctgggaatcc
     1861 aacgctggtg gttctttcac tgttactcta gacgaagtta atgaaagaat tggtaggggt
     1921 accatcttga ggttattctt gaaagatgac caattggagt acttggaaga aaagagaata
     1981 aaggaagtta tcaagagaca ttctgagttc gtggcctacc caatccaatt agtcgtcacc
     2041 aaggaagttg aaaaggaagt tccaattcca gaagaagaaa agaaagacga ggaaaagaag
     2101 gatgaggaaa agaaggatga agacgacaag aaaccaaaac tcgaggaagt cgatgaagaa
     2161 gaggaaaaga agccaaagac gaaaaaagtt aaagaagaag ttcaagagat agaagaacta
     2221 aacaagacta agcctttgtg gactagaaac ccatctgata tcactcaaga agaatacaat
     2281 gctttctata agtctatttc aaacgactgg gaagacccat tgtacgttaa gcatttctcc
     2341 gttgaaggtc aattggaatt tagagctatc ttattcattc caaagagagc accattcgac
     2401 ttgtttgaga gtaaaaagaa gaagaataat atcaagttgt acgttcgtcg tgttttcatc
     2461 actgatgaag ctgaagactt gattccagag tggttatctt tcgtcaaggg tgttgttgac
     2521 tctgaggatt taccattgaa tttgtccaga gaaatgttac aacaaaataa gatcatgaag
     2581 gttattagaa agaacattgt caaaaagttg attgaagcct tcaacgaaat tgctgaagac
     2641 tctgaacaat ttgaaaagtt ctactcggct ttctccaaaa atatcaagtt gggtgtacat
     2701 gaagataccc aaaacagggc tgctttggct aagttgttac gttacaactc taccaagtcc
     2761 gtagatgagt tgacttcctt aactgattac gttaccagaa tgccagaaca ccaaaagaac
     2821 atctactaca tcactggtga atctctaaag gctgtcgaaa agtctccatt tttggatgcc
     2881 ttgaaggcta aaaacttcga ggttttgttc ttgaccgacc caattgatga atacgccttc
     2941 actcaattga aggaattcga aggtaaaact ttggttgaca ttactaaaga tttcgaattg
     3001 gaagaaactg acgaagaaaa agctgaaaga gagaaggaga tcaaagaata tgaaccattg
     3061 accaaagctt tgaaagaaat tttgggtgac caagtggaga aagttgttgt ttcttacaaa
     3121 ttgttggatg ccccagctgc tatcagaact ggtcaatttg gttggtctgc taacatggaa
     3181 agaatcatga agtgtcaagc cttgagagac tcttccatgt cctcctacat gtcttccaag
     3241 aagactttcg aaatttctcc aaaatctcca attatcaagg aattgaaaaa gagagttgac
     3301 gaaggtggtg ctcaagacaa gactgtcaag gacttgacta agttattata tgaaactgct
     3361 ttgttgactt ccggcttcag tttggacgaa ccaacttcct ttgcatcaag aattaacaga
     3421 ttgatctctt taggcctgaa cattggtggt ggtactagtt ctgttaagga attggaagat
     3481 aagaacgaag aattgttgtc tagaattgct catgaaagaa acgaagttgc tagattgaag
     3541 aagttggttg gtgaaagaac cggtgatgag gatgaagaaa cagagactgc tccagaagca
     3601 tccaccgcag ctccggttga agaggttcca gctgacaccg aaatggaaga ggtagattaa
     3661 tggatccccc gggctgcagg aattcgatat caagcttatc gataccgtcg acctcgagtc
     3721 atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa
     3781 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta
     3841 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac
     3901 gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta
     3961 atttgcggcc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     4021 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     4081 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     4141 aacagttgcg cagcctgaat ggcgaatggc gcgacgcgcc ctgtagcggc gcattaagcg
     4201 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4261 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4321 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4381 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4441 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4501 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4561 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     4621 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4681 agatccgtcg agttcaagag aaaaaaaaag aaaaagcaaa aagaaaaaag gaaagcgcgc
     4741 ctcgttcaga atgacacgta tagaatgatg cattaccttg tcatcttcag tatcatactg
     4801 ttcgtataca tacttactga cattcatagg tatacatata tacacatgta tatatatcgt
     4861 atgctgcagc tttaaataat cggtgtcact acataagaac acctttggtg gagggaacat
     4921 cgttggtacc attgggcgag gtggcttctc ttatggcaac cgcaagagcc ttgaacgcac
     4981 tctcactacg gtgatgatca ttcttgcctc gcagacaatc aacgtggagg gtaattctgc
     5041 tagcctctgc aaagctttca agaaaatgcg ggatcatctc gcaagagaga tctcctactt
     5101 tctccctttg caaaccaagt tcgacaactg cgtacggcct gttcgaaaga tctaccaccg
     5161 ctctggaaag tgcctcatcc aaaggcgcaa atcctgatcc aaaccttttt actccacgcg
     5221 ccagtagggc ctctttaaaa gcttgaccga gagcaatccc gcagtcttca gtggtgtgat
     5281 ggtcgtctat gtgtaagtca ccaatgcact caacgattag cgaccagccg gaatgcttgg
     5341 ccagagcatg tatcatatgg tccagaaacc ctatacctgt gtggacgtta atcacttgcg
     5401 attgtgtggc ctgttctgct actgcttctg cctctttttc tgggaagatc gagtgctcta
     5461 tcgctagggg accacccttt aaagagatcg caatctgaat cttggtttca tttgtaatac
     5521 gctttactag ggctttctgc tctgtcatct ttgccttcgt ttatcttgcc tgctcatttt
     5581 ttagtatatt cttcgaagaa atcacattac tttatataat gtataattca ttatgtgata
     5641 atgccactcg ctaagaaaaa aaaagagtca tccgctaggt ggaaaaaaaa aaatgaaaat
     5701 cattaccgag gcataaaaaa atatagagtg tactagagga ggccaagagt aatagaaaaa
     5761 gaaaattgcg ggaaaggact gtgttatgac ttccctgact aatgccgtgt tcaaacgata
     5821 cctggcagtg actcctagcg ctcaccaagc tcttaaaacg ggaatttatg gtgcactctg
     5881 gcgcgccctg ggggaacaac ttcacagaat gttttgtcat attgtcgaag tggtcacaaa
     5941 acaagagaag ttccgccaat tataaaaagg gaacccgtat atttcagctt cacggatgat
     6001 ttccagggtg agagtactgt atatgggctt acgatagaag gccataaaaa tttcttgctt
     6061 ggcaacaaaa tagaagtgaa atcatgtcga ggctgctgtg tgggagaaca gcataaaata
     6121 tcacaaaaaa agaatctaaa acactgtgtt gcttgtccca gaaagggaat caagtatttt
     6181 tataaagatt ggagtggtaa aaatcgagta tgtgctagat gctatggaag atacaaattc
     6241 agcggtcatc actgtataaa ttgcaagtat gtaccagaag cacgtgaagt gaaaaaggca
     6301 aaagacaaag gcgaaaaatt gggcattacg cccgaaggtt tgccagttaa aggaccagag
     6361 tgtataaaat gtggcggaat cttacagtgg cctatgcggc cgctctagct agtggatcga
     6421 tccccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
     6481 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     6541 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     6601 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     6661 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     6721 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     6781 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     6841 gggtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     6901 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt
     6961 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     7021 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     7081 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     7141 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     7201 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     7261 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     7321 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     7381 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     7441 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     7501 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     7561 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     7621 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     7681 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     7741 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     7801 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     7861 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     7921 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     7981 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     8041 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     8101 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     8161 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     8221 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     8281 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     8341 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     8401 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     8461 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     8521 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     8581 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     8641 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     8701 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     8761 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     8821 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga
     8881 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     8941 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     9001 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     9061 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     9121 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     9181 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     9241 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     9301 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     9361 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc c
//
