LOCUS       sequence-456188-        6751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expressed human Hsp90 (G97D mutation) with a mOFP fluorescent 
            protein tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6751)
  AUTHORS   French JB, Zhao H, An S, Niessen S, Deng Y, Cravatt BF, Benkovic SJ
  TITLE     Hsp70/Hsp90 chaperone machinery is involved in the assembly of the 
            purinosome.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Feb 12;110(7):2528-33. doi: 
            10.1073/pnas.1300173110. Epub 2013 Jan 28.
  PUBMED    23359685
REFERENCE   2  (bases 1 to 6751)
  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: 
            "10.1073/pnas.1300173110"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2013-02-12- 12"; volume: "110"; issue: "7"; pages: "2528-33"
COMMENT     Sequence Label: pHSP90(G97D)-mOFP
FEATURES             Location/Qualifiers
     source          1..6751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(172..760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(252..271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1089..1136
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1161..1180
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1368..2162)
                     /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(1368..2162)
                     /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(1479..1498)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2089..2108
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2197..2554)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2211..2346
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2363..2554
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2511..2531
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2556..2660)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2687..3142
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2774..2793)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2984..3005
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3149..3270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3160..3179)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3214..3233
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3285..3995)
                     /codon_start=1
                     /product="orange monomeric derivative of DsRed fluorescent 
                     protein (Shanet et al., 2004)"
                     /label=mOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3563..3582)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      3992..4001
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6241..6444)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6270..6290)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6445..6748
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactaatg
       61 catggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      121 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
      181 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      241 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      301 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      361 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      421 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      481 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
      541 tagcagagcg aggtatgtaa gcggtgctac agagttcttg aagtggtggc ctaactacgg
      601 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
      661 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
      721 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
      781 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa tggattttgg tcatgagatt
      841 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
      901 aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag
      961 ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta
     1021 ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc
     1081 gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat
     1141 agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg
     1201 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     1261 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     1321 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     1381 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     1441 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     1501 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     1561 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     1621 ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     1681 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     1741 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     1801 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     1861 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     1921 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct
     1981 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     2041 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     2101 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     2161 atgcgaaacg atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa
     2221 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     2281 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     2341 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     2401 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     2461 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     2521 acacacattc cacagctggt tctttccgcc tcaggactct tcctttttca atattattga
     2581 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     2641 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     2701 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2761 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2821 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2881 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2941 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3001 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3061 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3121 aaaatattaa cgcttacaat ttacgcctta agatacattg atgagtttgg acaaaccaca
     3181 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3241 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attcttactt gtacagctcg
     3301 tccatgccgc cggtggagtg gcggccctcg gcgcgttcgt actgttccac gatggtgtag
     3361 tcctcgttgt gggaggtgat gtccaacttg atgccgacga tgtaggcgcc gggcagctgc
     3421 acgggcttct tggccttgta ggtggtcttg acctcggagg tgtagtggcc gccgtccttc
     3481 agcttcagcc tcatcttgat ctcgcccttc agggcgccgt cctcggggta catccgctcg
     3541 gaggaggcct cccagcccat ggtcttcttc tgcattacgg ggccgtcgga ggggaagttg
     3601 gtgccgcgca gcttcacctt gtagatgaac tcgccgtcct gcagggagga gtcctgggtc
     3661 acggtcacca cgccgccgtc ctcgaagttc atcacgcgct cccacttgaa gccctcgggg
     3721 aaggacagct tgaagtagtc ggggatgtcg gcggggtgct tcacgtaggc cttggagccg
     3781 taggtgaact gaggggacag gatgtcccag gcgaagggca gggggccacc cttggtcacc
     3841 ttcagcttag cggtctgaaa gccctcgtag gggcggccct cgccctcgcc ctcgatctcg
     3901 aactcgtggc cgttcacgga gccctccatg cgcaccttga agcgcatgaa ctccttgatg
     3961 atggccatgt tattctcctc gcccttgctc accatggtgg cgggatccag agattcgtct
     4021 acttcttcca tgcgtgatgt gtcgtcatct ccttcaaggg gtggcatttc ttcagttaca
     4081 gcagcactgg tatcatcagc agtagggtca tcttcatcaa tacccagacc aagtttgatc
     4141 atcctgtaga tcctgttagc atgtgtctgg ggatcttcca gactgaagcc agaagacagg
     4201 agcgcagttt cataaagcaa gatgaccaga tccttcacag acttgtcgtt cttatcagcc
     4261 tctgcctttt gccttaaggt ctcaataatg gaatggtcag ggtttatctc caggtgtttc
     4321 tttgctgcca tgtaacccat tgttgagttg tctcttaggg cttgagcttt catgattctc
     4381 tccatgtttg ctgtccagcc atatgtgctt gtgacaatac agcatggaga tgtcaccaat
     4441 cggtttgaca caaccacctt ttcaactttt ttctccaata tgtctttcat gattttgcag
     4501 aggttctcaa actttgtttt tttctcttcc tgcttctttt tctcttcttc atcctctgga
     4561 agttccaggc cttctttggt gactgacact aaagtcttcc cctcaaattc cttcagctgt
     4621 tggacacagt actcatcaat gggctcaatc atatagatca cttctaagcc atgtttccga
     4681 agacgttcca caaaggctga gttagctacc tggtccttgg tctcacctgt gatataatag
     4741 atatgtttct ggttctcctt cattctggtg cagtagtcct tgagagaaac catctcatca
     4801 ccagaggcag atgtgtagta ccttaacagc tctgaaagct tcttccgatt ttgagagtct
     4861 tcgtgtattc caagctttat gtttttagag aactgctcat agaatttctt gtagttctct
     4921 ttatcttccg ccagttcagt aaagagttct aagcattttt tgaccaaatt cttcctgata
     4981 actttcaaaa ttttgctttg ttgcaacatc tcacgggata tgtttagagg gagatcctcc
     5041 gagtctacca cccctctaat gaagttcaga tattcaggga ttagctcctc acagttatcc
     5101 atgatgaaaa ctctgcgtac atacaatttg atattgttct ttttctttct gttttcaaac
     5161 agatcaaaag gagcacgtcg tgggacaaat agaagggctc tgaattccaa ctgtccttca
     5221 actgaaaaat gcttcactgc caagtgatct tcccagtcat tggtcaagct cttatagaat
     5281 tctccgtact cctcattagt aatatcgtcg ggatttctgg tccagatggg ctttgttttg
     5341 ttgagctctt cttgatcgat gtacttttcc ttaatcttct tcttcttctt cttgtcacca
     5401 tccttctttt cttcttcctc atcagaacca acatcttcaa tttcaggttt gtcttccgac
     5461 tctttctctt ctttttcttt ttcttcttct ttgtcttcct tttcttcagc ctcatcatcg
     5521 cttacttctt tatcacgttc cttctccaca aaaagagtaa tgggatatcc aataaactga
     5581 gaatgtttct tcacaatctc ctttattctt cgttcctcca agtactcagt ttggtcttct
     5641 ttcaggtgta ggataacttt tgttccacga cccataggtt cacctgtgtc tgtcctcact
     5701 gtgaatgatc cccctgctga ggactcccaa gcgtactgct catcatcgtt atgtttggtg
     5761 atcacagtta ctttctcagc aaccaaataa gcagaataaa aaccaacacc gaactggcca
     5821 atcatagaga tatctgcacc agcctgcaaa gcttccatga acgctttggt cccagacttg
     5881 gcgatagtac caaggttatt gatcaagtca gccttggtca tatcaattcc agtatccaca
     5941 atagtgagag ttcgatcttg tttgttcggt ataaggttaa tatgcagctc tttcccagag
     6001 tctaatttac tgggatctgt caagctttca taccggattt tgtccaatgc atctgatgaa
     6061 tttgaaatga gctctctcag aaagatctct ttgttcgagt agaaagtatt gatgatcaat
     6121 gacatcaact gggcaatttc tgcctgaaag gcgaacgtct caacctcctc ctcctccatc
     6181 ggttggtctt gggtctgggt ttcctcaggc atgctagcgg atctgacggt tcactaaacc
     6241 agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc
     6301 ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt
     6361 gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt
     6421 gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg
     6481 ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc
     6541 gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg
     6601 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     6661 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     6721 cgggccattt accgtaagtt atgtaacgcg g
//
