LOCUS       sequence-54824-e        7835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7835)
  AUTHORS   Wayne N, Bolon DN
  TITLE     Dimerization of Hsp90 is required for in vivo function. Design and 
            analysis of monomers and dimers.
  JOURNAL   J Biol Chem. 2007 Nov 30;282(48):35386-95. Epub 2007 Oct 1.
  PUBMED    17908693
REFERENCE   2  (bases 1 to 7835)
  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: "J Biol 
            Chem."; date: "2007-11-30"; volume: "282(48)"; pages: "35386-95. 
            Epub 2007 Oct 1"
COMMENT     Sequence Label: pRS414-GPD-6xHis-Hsp82
FEATURES             Location/Qualifiers
     source          1..7835
                     /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             3470..3487
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5666..5682)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5667..5683)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5685..5932
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5701..5719)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5950..5969)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5951..5969)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5979..5996)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5979..5995)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5988..6010)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6140..6595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6227..6246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6437..6458
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6694..7368)
                     /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(7369..7649)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7666..7685)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7785..7807
                     /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 ttagtctttt ttttagtttt
     3421 aaaacaccag aacttagttt cgacggattc tagaactagt atgggtggtc accaccatca
     3481 ccaccacggt ggtcatatgg ctagtgaaac ttttgaattt caagctgaaa ttactcagtt
     3541 gatgagtttg atcatcaaca ccgtctattc taacaaggaa attttcttga gagaactgat
     3601 atctaatgcc tcggatgcgt tggataaaat tagatacaaa tctttgtctg atccaaagca
     3661 attggaaaca gaaccagatc tctttattag aatcactcca aagccagagc aaaaagtttt
     3721 ggaaatcaga gattctggta ttggtatgac caaggctgaa ttgattaata acttgggtac
     3781 cattgccaag tctggtacca aagccttcat ggaagctcta tctgctggtg ccgatgtatc
     3841 catgattggt caattcggtg ttggttttta ctctttattc ttagttgccg acagagttca
     3901 ggttatttca aagagcaacg acgacgaaca atacatctgg gaatccaacg ctggtggttc
     3961 tttcactgtt actctagacg aagttaatga aagaattggt aggggtacca tcttgaggtt
     4021 attcttgaaa gatgaccaat tggagtactt ggaagaaaag agaataaagg aagttatcaa
     4081 gagacattct gagttcgtgg cctacccaat ccaattagtc gtcaccaagg aagttgaaaa
     4141 ggaagttcca attccagaag aagaaaagaa agacgaggaa aagaaggatg aggaaaagaa
     4201 ggatgaagac gacaagaaac caaaactcga ggaagtcgat gaagaagagg aaaagaagcc
     4261 aaagacgaaa aaagttaaag aagaagttca agagatagaa gaactaaaca agactaagcc
     4321 tttgtggact agaaacccat ctgatatcac tcaagaagaa tacaatgctt tctataagtc
     4381 tatttcaaac gactgggaag acccattgta cgttaagcat ttctccgttg aaggtcaatt
     4441 ggaatttaga gctatcttat tcattccaaa gagagcacca ttcgacttgt ttgagagtaa
     4501 aaagaagaag aataatatca agttgtacgt tcgtcgtgtt ttcatcactg atgaagctga
     4561 agacttgatt ccagagtggt tatctttcgt caagggtgtt gttgactctg aggatttacc
     4621 attgaatttg tccagagaaa tgttacaaca aaataagatc atgaaggtta ttagaaagaa
     4681 cattgtcaaa aagttgattg aagccttcaa cgaaattgct gaagactctg aacaatttga
     4741 aaagttctac tcggctttct ccaaaaatat caagttgggt gtacatgaag atacccaaaa
     4801 cagggctgct ttggctaagt tgttacgtta caactctacc aagtccgtag atgagttgac
     4861 ttccttaact gattacgtta ccagaatgcc agaacaccaa aagaacatct actacatcac
     4921 tggtgaatct ctaaaggctg tcgaaaagtc tccatttttg gatgccttga aggctaaaaa
     4981 cttcgaggtt ttgttcttga ccgacccaat tgatgaatac gccttcactc aattgaagga
     5041 attcgaaggt aaaactttgg ttgacattac taaagatttc gaattggaag aaactgacga
     5101 agaaaaagct gaaagagaga aggagatcaa agaatatgaa ccattgacca aagctttgaa
     5161 agaaattttg ggtgaccaag tggagaaagt tgttgtttct tacaaattgt tggatgcccc
     5221 agctgctatc agaactggtc aatttggttg gtctgctaac atggaaagaa tcatgaaggc
     5281 tcaagccttg agagactctt ccatgtcctc ctacatgtct tccaagaaga ctttcgaaat
     5341 ttctccaaaa tctccaatta tcaaggaatt gaaaaagaga gttgacgaag gtggtgctca
     5401 agacaagact gtcaaggact tgactaagtt attatatgaa actgctttgt tgacttccgg
     5461 cttcagtttg gacgaaccaa cttcctttgc atcaagaatt aacagattga tctctttagg
     5521 cctgaacatt gatgaggatg aagaaacaga gactgctcca gaagcatcca ccgcagctcc
     5581 ggttgaagag gttccagctg acaccgaaat ggaagaggta gattaatgga tcccccgggc
     5641 tgcaggaatt cgatatcaag cttatcgata ccgtcgacct cgagtcatgt aattagttat
     5701 gtcacgctta cattcacgcc ctccccccac atccgctcta accgaaaagg aaggagttag
     5761 acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt
     5821 atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat
     5881 actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt gcggccggta
     5941 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     6001 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     6061 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     6121 ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     6181 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     6241 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     6301 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     6361 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     6421 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     6481 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     6541 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttcctga
     6601 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggc aagtgcacaa
     6661 acaatactta aataaatact actcagtaat aacctatttc ttagcatttt tgacgaaatt
     6721 tgctattttg ttagagtctt ttacaccatt tgtctccaca cctccgctta catcaacacc
     6781 aataacgcca tttaatctaa gcgcatcacc aacattttct ggcgtcagtc caccagctaa
     6841 cataaaatgt aagctttcgg ggctctcttg ccttccaacc cagtcagaaa tcgagttcca
     6901 atccaaaagt tcacctgtcc cacctgcttc tgaatcaaac aagggaataa acgaatgagg
     6961 tttctgtgaa gctgcactga gtagtatgtt gcagtctttt ggaaatacga gtcttttaat
     7021 aactggcaaa ccgaggaact cttggtattc ttgccacgac tcatctccat gcagttggac
     7081 gatatcaatg ccgtaatcat tgaccagagc caaaacatcc tccttaggtt gattacgaaa
     7141 cacgccaacc aagtatttcg gagtgcctga actattttta tatgctttta caagacttga
     7201 aattttcctt gcaataaccg ggtcaattgt tctctttcta ttgggcacac atataatacc
     7261 cagcaagtca gcatcggaat ctagagcaca ttctgcggcc tctgtgctct gcaagccgca
     7321 aactttcacc aatggaccag aactacctgt gaaattaata acagacatac tccaagctgc
     7381 ctttgtgtgc ttaatcacgt atactcacgt gctcaatagt caccaatgcc ctccctcttg
     7441 gccctctcct tttctttttt cgaccgaatt aattcttaat cggcaaaaaa agaaaagctc
     7501 cggatcaaga ttgtacgtaa ggtgacaagc tatttttcaa taaagaatat cttccactac
     7561 tgccatctgg cgtcataact gcaaagtaca catatattac gatgctgtct attaaatgct
     7621 tcctatatta tatatatagt aatgtcgttt atggtgcact ctcagtacaa tctgctctga
     7681 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     7741 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     7801 tcagaggttt tcaccgtcat caccgaaacg cgcga
//
