LOCUS       sequence-104527-        7956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast plasmid with Hsp82 heterodimer construct..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7956)
  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 7956)
  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: p414GPD_Hsp82viiB
FEATURES             Location/Qualifiers
     source          1..7956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /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(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2235..2254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2658..2663,2664..2681,2682..2688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /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)."
     primer_bind     2701..2723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2720..2736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2755..2775
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2800..3443
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3337..3360
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             3471..3488
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5787..5803)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5788..5804)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5806..6053
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5822..5840)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6071..6090)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6072..6090)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6100..6117)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6100..6116)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6109..6131)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6261..6716
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6348..6367)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6558..6579
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6815..7489)
                     /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"
     promoter        complement(7490..7770)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7787..7806)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7906..7928
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     2761 aaccctcact aaagggaaca aaagctggag ctcagtttat cattatcaat actcgccatt
     2821 tcaaagaata cgtaaataat taatagtagt gattttccta actttattta gtcaaaaaat
     2881 tagcctttta attctgctgt aacccgtaca tgcccaaaat agggggcggg ttacacagaa
     2941 tatataacat cgtaggtgtc tgggtgaaca gtttattcct ggcatccact aaatataatg
     3001 gagcccgctt tttaagctgg catccagaaa aaaaaagaat cccagcacca aaatattgtt
     3061 ttcttcacca accatcagtt cataggtcca ttctcttagc gcaactacag agaacagggg
     3121 cacaaacagg caaaaaacgg gcacaacctc aatggagtga tgcaacctgc ctggagtaaa
     3181 tgatgacaca aggcaattga cccacgcatg tatctatctc attttcttac accttctatt
     3241 accttctgct ctctctgatt tggaaaaagc tgaaaaaaaa ggttgaaacc agttccctga
     3301 aattattccc ctacttgact aataagtata taaagacggt aggtattgat tgtaattctg
     3361 taaatctatt tcttaaactt cttaaattct acttttatag ttagtcattt ttttagtttt
     3421 aaaacaccag aacttagttt cgagaaaaat agaatactag tatgggtggt caccaccatc
     3481 accaccacgg tggtcatatg gctagtgaaa cttttgaatt tcaagctgaa attactcagt
     3541 tgatgagttt gatcatcaac accgtctatt ctaacaagga aattttcttg agagaactga
     3601 tatctaatgc ctcggatgcg ttggataaaa ttagatacaa atctttgtct gatccaaagc
     3661 aattggaaac agaaccagat ctctttatta gaatcactcc aaagccagag caaaaagttt
     3721 tggaaatcag agattctggt attggtatga ccaaggctga attgattaat aacttgggta
     3781 ccattgccaa gtctggtacc aaagccttca tggaagctct atctgctggt gccgatgtat
     3841 ccatgattgg tcaattcggt gttggttttt actctttatt cttagttgcc gacagagttc
     3901 aggttatttc aaagagcaac gacgacgaac aatacatctg ggaatccaac gctggtggtt
     3961 ctttcactgt tactctagac gaagttaatg aaagaattgg taggggtacc atcttgaggt
     4021 tattcttgaa agatgaccaa ttggagtact tggaagaaaa gagaataaag gaagttatca
     4081 agagacattc tgagttcgtg gcctacccaa tccaattagt cgtcaccaag gaagttgaaa
     4141 aggaagttcc aattccagaa gaagaaaaga aagacgagga aaagaaggat gaggaaaaga
     4201 aggatgaaga cgacaagaaa ccaaaactcg aggaagtcga tgaagaagag gaaaagaagc
     4261 caaagacgaa aaaagttaaa gaagaagttc aagagataga agaactaaac aagactaagc
     4321 ctttgtggac tagaaaccca tctgatatca ctcaagaaga atacaatgct ttctataagt
     4381 ctatttcaaa cgactgggaa gacccattgt acgttaagca tttctccgtt gaaggtcaat
     4441 tggaatttag agctatctta ttcattccaa agagagcacc attcgacttg tttgagagta
     4501 aaaagaagaa gaataatatc aagttgtacg ttcgtcgtgt tttcatcact gatgaagctg
     4561 aagacttgat tccagagtgg ttatctttcg tcaagggtgt tgttgactct gaggatttac
     4621 cattgaattt gtccagagaa atgttacaac aaaataagat catgaaggtt attagaaaga
     4681 acattgtcaa aaagttgatt gaagccttca acgaaattgc tgaagactct gaacaatttg
     4741 aaaagttcta ctcggctttc tccaaaaata tcaagttggg tgtacatgaa gatacccaaa
     4801 acagggctgc tttggctaag ttgttacgtt acaactctac caagtccgta gatgagttga
     4861 cttccttaac tgattacgtt accagaatgc cagaacacca aaagaacatc tactacatca
     4921 ctggtgaatc tctaaaggct gtcgaaaagt ctccattttt ggatgccttg aaggctaaaa
     4981 acttcgaggt tttgttcttg accgacccaa ttgatgaata cgccttcact caattgaagg
     5041 aattcgaagg taaaactttg gttgacatta ctaaagattt cgaattggaa gaaactgacg
     5101 aagaaaaagc tgaaagagag aaggagatca aagaatatga accattgacc aaagctttga
     5161 aagaaatttt gggtgaccaa gtggagaaag ttgttgtttc ttacaaattg ttggatgccc
     5221 cagctgctat cagaactggt caatttggtt ggtctgctaa catggaaaga atcatgaagt
     5281 gtcaagcctt gagagactct tccatgtcct cctacatgtc ttccaagaag actttcgaaa
     5341 tttctccaaa atctccaatt atcaaggaat tgaaaaagag agttgacgaa ggtggtgctc
     5401 aagacaagac tgtcaaggac ttgactaagt tattatatga aactgctttg ttgacttccg
     5461 gcttcagttt ggacgaacca acttcctttg catcaagaat taacagattg atctctttag
     5521 gcctgaacat tggtggtggt actagttctg ttaaggaatt ggaagataag aacgaagaat
     5581 tgttgtctga attggctcat ttgaagaacg aagttgctag attgaagaag ttggttggtg
     5641 aaagaaccgg tgatgaggat gaagaaacag agactgctcc agaagcatcc accgcagctc
     5701 cggttgaaga ggttccagct gacaccgaaa tggaagaggt agattaatgg atcccccggg
     5761 ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc tcgagtcatg taattagtta
     5821 tgtcacgctt acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta
     5881 gacaacctga agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt
     5941 tatttatatt tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta
     6001 tactgaaaac cttgcttgag aaggttttgg gacgctcgaa ggctttaatt tgcggccggt
     6061 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     6121 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     6181 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     6241 cctgaatggc gaatggcgcg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     6301 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     6361 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6421 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     6481 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     6541 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     6601 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     6661 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caatttcctg
     6721 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatagg caagtgcaca
     6781 aacaatactt aaataaatac tactcagtaa taacctattt cttagcattt ttgacgaaat
     6841 ttgctatttt gttagagtct tttacaccat ttgtctccac acctccgctt acatcaacac
     6901 caataacgcc atttaatcta agcgcatcac caacattttc tggcgtcagt ccaccagcta
     6961 acataaaatg taagctttcg gggctctctt gccttccaac ccagtcagaa atcgagttcc
     7021 aatccaaaag ttcacctgtc ccacctgctt ctgaatcaaa caagggaata aacgaatgag
     7081 gtttctgtga agctgcactg agtagtatgt tgcagtcttt tggaaatacg agtcttttaa
     7141 taactggcaa accgaggaac tcttggtatt cttgccacga ctcatctcca tgcagttgga
     7201 cgatatcaat gccgtaatca ttgaccagag ccaaaacatc ctccttaggt tgattacgaa
     7261 acacgccaac caagtatttc ggagtgcctg aactattttt atatgctttt acaagacttg
     7321 aaattttcct tgcaataacc gggtcaattg ttctctttct attgggcaca catataatac
     7381 ccagcaagtc agcatcggaa tctagagcac attctgcggc ctctgtgctc tgcaagccgc
     7441 aaactttcac caatggacca gaactacctg tgaaattaat aacagacata ctccaagctg
     7501 cctttgtgtg cttaatcacg tatactcacg tgctcaatag tcaccaatgc cctccctctt
     7561 ggccctctcc ttttcttttt tcgaccgaat taattcttaa tcggcaaaaa aagaaaagct
     7621 ccggatcaag attgtacgta aggtgacaag ctatttttca ataaagaata tcttccacta
     7681 ctgccatctg gcgtcataac tgcaaagtac acatatatta cgatgctgtc tattaaatgc
     7741 ttcctatatt atatatatag taatgtcgtt tatggtgcac tctcagtaca atctgctctg
     7801 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     7861 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     7921 gtcagaggtt ttcaccgtca tcaccgaaac gcgcga
//
