LOCUS       sequence-207848-        6887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for EGFP fusion to human Hsp90 beta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6887)
  AUTHORS   Picard D, Suslova E, Briand PA
  TITLE     2-color photobleaching experiments reveal distinct intracellular 
            dynamics of two components of the Hsp90 complex.
  JOURNAL   Exp Cell Res. 2006 Nov 15. 312(19):3949-58.
  PUBMED    17010336
REFERENCE   2  (bases 1 to 6887)
  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: "Exp Cell 
            Res. 2006 Nov 15. 312(19):3949-58."
COMMENT     Sequence Label: pEGFP.90 beta
FEATURES             Location/Qualifiers
     source          1..6887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        99..402
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        403..606
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     557..577
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      645..654
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(651..653,654..656,657..1367)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(651..653,654..656,657..1367)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(696..717)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(957..976)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1304..1325
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3713..3834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3750..3769)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3804..3823
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3841..4296)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3978..3999)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4190..4209
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4323..4427
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4429..4786
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4429..4620
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4452..4472)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4637..4772
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4821..5615
                     /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             4821..5615
                     /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(4875..4894)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5485..5504
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5803..5822)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5847..5894
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6223..6811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6712..6731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
       61 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      121 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      181 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      241 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      301 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      361 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      421 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      481 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      541 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      601 agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc atggtgagca
      661 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      721 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      781 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      841 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
      901 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      961 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1021 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1081 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1141 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1201 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1261 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1321 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc ggactcagat
     1381 ctggaggtat gcctgaggaa gtgcaccatg gagaggagga ggtggagact tttgcctttc
     1441 aggcagaaat tgcccaactc atgtccctca tcatcaatac cttctattcc aacaaggaga
     1501 ttttccttcg ggagttgatc tctaatgctt ctgatgcctt ggacaagatt cgctatgaga
     1561 gcctgacaga cccttcgaag ttggacagtg gtaaagagct gaaaattgac atcatcccca
     1621 accctcagga acgtaccctg actttggtag acacaggcat tggcatgacc aaagctgatc
     1681 tcataaataa tttgggaacc attgccaagt ctggtactaa agcattcatg gaggctcttc
     1741 aggctggtgc agacatctcc atgattgggc agtttggtgt tggcttttat tctgcctact
     1801 tggtggcaga gaaagtggtt gtgatcacaa agcacaacga tgatgaacag tatgcttggg
     1861 agtcttctgc tggaggttcc ttcactgtgc gtgctgacca tggtgagccc attggcaggg
     1921 gtaccaaagt gatcctccat cttaaagaag atcagacaga gtacctagaa gagaggcggg
     1981 tcaaagaagt agtgaagaag cattctcagt tcataggcta tcccatcacc ctttatttgg
     2041 agaaggaacg agagaaggaa attagtgatg atgaggcaga ggaagagaaa ggtgagaaag
     2101 aagaggaaga taaagatgat gaagaaaagc ccaagatcga agatgtgggt tcagatgagg
     2161 aggatgacag cggtaaggat aagaagaaga aaactaagaa gatcaaagag aaatacattg
     2221 atcaggaaga actaaacaag accaagccta tttggaccag aaaccctgat gacatcaccc
     2281 aagaggagta tggagaattc tacaagagcc tcactaatga ctgggaagac cacttggcag
     2341 tcaagcactt ttctgtagaa ggtcagttgg aattcagggc attgctattt attcctcgtc
     2401 gggctccctt tgaccttttt gagaacaaga agaaaaagaa caacatcaaa ctctatgtcc
     2461 gccgtgtgtt catcatggac agctgtgatg agttgatacc agagtatctc aattttatcc
     2521 gtggtgtggt tgactctgag gatctgcccc tgaacatctc ccgagaaatg ctccagcaga
     2581 gcaaaatctt gaaagtcatt cgcaaaaaca ttgttaagaa gtgccttgag ctcttctctg
     2641 agctggcaga agacaaggag aattacaaga aattctatga ggcattctct aaaaatctca
     2701 agcttggaat ccacgaagac tccactaacc gccgccgcct gtctgagctg ctgcgctatc
     2761 atacctccca gtctggagat gagatgacat ctctgtcaga gtatgtttct cgcatgaagg
     2821 agacacagaa gtccatctat tacatcactg gtgagagcaa agagcaggtg gccaactcag
     2881 cttttgtgga gcgagtgcgg aaacggggct tcgaggtggt atatatgacc gagcccattg
     2941 acgagtactg tgtgcagcag ctcaaggaat ttgatgggaa gagcctggtc tcagttacca
     3001 aggagggtct ggagctgcct gaggatgagg aggagaagaa gaagatggaa gagagcaagg
     3061 caaagtttga gaacctctgc aagctcatga aagaaatctt agataagaag gttgagaagg
     3121 tgacaatctc caatagactt gtgtcttcac cttgctgcat tgtgaccagc acctacggct
     3181 ggacagccaa tatggagcgg atcatgaaag cccaggcact tcgggacaac tccaccatgg
     3241 gctatatgat ggccaaaaag cacctggaga tcaaccctga ccaccccatt gtggagacgc
     3301 tgcggcagaa ggctgaggcc gacaagaatg ataaggcagt taaggacctg gtggtgctgc
     3361 tgtttgaaac cgccctgcta tcttctggct tttcccttga ggatccccag acccactcca
     3421 accgcatcta tcgcatgatc aagctaggtc taggtattga tgaagatgaa gtggcagcag
     3481 aggaacccaa tgctgcagtt cctgatgaga tcccccctct cgagggcgat gaggatgcgt
     3541 ctcgcatgga agaagtcgat tagtcgacgg taccgcgggc ccgggatcca ccggatctag
     3601 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3661 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3721 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3781 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     3841 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3901 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3961 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4021 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4081 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4141 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4201 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4261 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     4321 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4381 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     4441 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4501 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4561 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4621 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4681 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4741 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     4801 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4861 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4921 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4981 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     5041 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5101 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5161 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5221 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5281 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5341 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5401 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5461 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5521 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5581 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5641 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5701 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5761 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     5821 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     5881 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5941 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     6001 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     6061 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6121 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6181 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6241 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6301 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6361 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6421 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6481 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6541 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6601 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6661 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6721 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6781 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6841 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttat
//
