LOCUS       sequence-217627-        5994 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Please note this plasmid contains GST-Hsp90 residues 273-617..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5994)
  AUTHORS   Fontana J, Fulton D, Chen Y, Fairchild TA, McCabe TJ, Fujita N, 
            Tsuruo T, Sessa WC
  TITLE     Domain mapping studies reveal that the M domain of hsp90 serves as a
            molecular scaffold to regulate Akt-dependent phosphorylation of 
            endothelial nitric oxide synthase and NO release.
  JOURNAL   Circ Res. 2002 May 3. 90(8):866-73.
  PUBMED    11988487
REFERENCE   2  (bases 1 to 5994)
  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: "Circ Res. 
            2002 May 3. 90(8):866-73."
COMMENT     Sequence Label: GST-Hsp90 M(272-617)
FEATURES             Location/Qualifiers
     source          1..5994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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)."
     CDS             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             736..753
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1862..1884)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2029..2047)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2115..2219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2220..2288,2289..3080)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2438..2457)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3251..3839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3740..3759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3993..4010
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4083..4160
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4161..5243
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4180..4199)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5292..5297,5298..5315,5316..5322)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5330..5346
                     /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     5354..5370
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5354..5370
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5366..5893
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5386..5403)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5386..5402)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5395..5417)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaat cggatctggt tccgcgtgga tccgacaaga agaagaagaa gaagattaag
      781 gaaaagtaca tcgatcaaga agagctcaac aaaacaaagc ccatctggac cagaaatccc
      841 gacgatatta ctaatgagga gtacggagaa ttctataaga gcttgaccaa tgactgggaa
      901 gatcacttgg cagtgaagca tttttcagtt gaaggacagt tggaattcag agcccttcta
      961 tttgtcccac gacgtgctcc ttttgatctg tttgaaaaca gaaagaaaaa gaacaatatc
     1021 aaattgtatg tacgcagagt tttcatcatg gataactgtg aggagctaat ccctgaatat
     1081 ctgaacttca ttagaggggt ggtagactcg gaggatctcc ctctaaacat atcccgtgag
     1141 atgttgcaac aaagcaaaat tttgaaagtt atcaggaaga atttggtcaa aaaatgctta
     1201 gaactcttta ctgaactggc ggaagataaa gagaactaca agaaattcta tgagcagttc
     1261 tctaaaaaca taaagcttgg aatacacgaa gactctcaaa atcggaagaa gctttcagag
     1321 ctgttaaggt actacacatc tgcctctggt gatgagatgg tttctctcaa ggactactgc
     1381 accagaatga aggagaacca gaaacatatc tattatatca caggtgagac caaggaccag
     1441 gtagctaact cagcctttgt ggaacgtctt cggaaacatg gcttagaagt gatctatatg
     1501 attgagccca ttgatgagta ctgtgtccaa cagctgaagg aatttgaggg gaagacttta
     1561 gtgtcagtca ccaaagaagg cctggaactt ccagaggatg aagaagagaa aaagaagcag
     1621 gaagagaaaa aaacaaagtt tgagaacctc tgcaaaatca tgaaagacat attggagaaa
     1681 aaagttgaaa aggtggttgt gtcaaaccga ttggtgacat ctccatgctg tattgtcaca
     1741 agcacatatg gctggacagc aaacatggag agaatcatga aagctcaata ggtcgactcg
     1801 agcggccgca tcgtgactga ctgacgatct gcctcgcgcg tttcggtgat gacggtgaaa
     1861 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     1921 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     1981 cccagtcacg tagcgatagc ggagtgtata attcttgaag acgaaagggc ctcgtgatac
     2041 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     2101 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2161 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2221 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2281 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2341 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2401 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2461 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2521 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2581 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2641 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2701 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2761 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2821 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2881 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     2941 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3001 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3061 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3121 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3181 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3241 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3301 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3361 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3421 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3481 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3541 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3601 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3661 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3721 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3781 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3841 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3901 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3961 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     4021 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt
     4081 ccgacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
     4141 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     4201 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     4261 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     4321 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
     4381 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
     4441 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
     4501 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
     4561 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
     4621 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
     4681 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
     4741 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
     4801 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
     4861 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     4921 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
     4981 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     5041 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     5101 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     5161 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     5221 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     5281 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     5341 taacaatttc acacaggaaa cagctatgac catgattacg gattcactgg ccgtcgtttt
     5401 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     5461 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     5521 gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg gcaccagaag cggtgccgga
     5581 aagctggctg gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct caaactggca
     5641 gatgcacggt tacgatgcgc ccatctacac caacgtgacc tatcccatta cggtcaatcc
     5701 gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta atgttgatga
     5761 aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttggaa ttagcttatc
     5821 gactgcacgg tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct
     5881 gtgcaggtcg taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata
     5941 atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga gctg
//
