LOCUS       sequence-204033-        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        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3675..3796
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3712..3731)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3766..3785
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3803..4258)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3940..3961)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4152..4171
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4285..4389
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4391..4748
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4391..4582
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4414..4434)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4599..4734
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4783..5577
                     /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             4783..5577
                     /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(4837..4856)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5447..5466
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5765..5784)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5809..5856
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6185..6773
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6674..6693
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctggaggt atgcctgagg aagtgcacca tggagaggag
     1381 gaggtggaga cttttgcctt tcaggcagaa attgcccaac tcatgtccct catcatcaat
     1441 accttctatt ccaacaagga gattttcctt cgggagttga tctctaatgc ttctgatgcc
     1501 ttggacaaga ttcgctatga gagcctgaca gacccttcga agttggacag tggtaaagag
     1561 ctgaaaattg acatcatccc caaccctcag gaacgtaccc tgactttggt agacacaggc
     1621 attggcatga ccaaagctga tctcataaat aatttgggaa ccattgccaa gtctggtact
     1681 aaagcattca tggaggctct tcaggctggt gcagacatct ccatgattgg gcagtttggt
     1741 gttggctttt attctgccta cttggtggca gagaaagtgg ttgtgatcac aaagcacaac
     1801 gatgatgaac agtatgcttg ggagtcttct gctggaggtt ccttcactgt gcgtgctgac
     1861 catggtgagc ccattggcag gggtaccaaa gtgatcctcc atcttaaaga agatcagaca
     1921 gagtacctag aagagaggcg ggtcaaagaa gtagtgaaga agcattctca gttcataggc
     1981 tatcccatca ccctttattt ggagaaggaa cgagagaagg aaattagtga tgatgaggca
     2041 gaggaagaga aaggtgagaa agaagaggaa gataaagatg atgaagaaaa acccaagatc
     2101 gaagatgtgg gttcagatga ggaggatgac agcggtaagg ataagaagaa gaaaactaag
     2161 aagatcaaag agaaatacat tgatcaggaa gaactaaaca agaccaagcc tatttggacc
     2221 agaaaccctg atgacatcac ccaagaggag tatggagaat tctacaagag cctcactaat
     2281 gactgggaag accacttggc agtcaagcac ttttctgtag aaggtcagtt ggaattcagg
     2341 gcattgctat ttattcctcg tcgggctccc tttgaccttt ttgagaacaa gaagaaaaag
     2401 aacaacatca aactctatgt ccgccgtgtg ttcatcatgg acagctgtga tgagttgata
     2461 ccagagtatc tcaattttat ccgtggtgtg gttgactctg aggatctgcc cctgaacatc
     2521 tcccgagaaa tgctccagca gagcaaaatc ttgaaagtca ttcgcaaaaa cattgttaag
     2581 aagtgccttg agctcttctc tgagctggca gaagacaagg agaattacaa gaaattctat
     2641 gaggcattct ctaaaaatct caagcttgga atccacgaag actccactaa ccgccgccgc
     2701 ctgtctgagc tgctgcgcta tcatacctcc cagtctggag atgagatgac atctctgtca
     2761 gagtatgttt ctcgcatgaa ggagacacag aagtccatct attacatcac tggtgagagc
     2821 aaagagcagg tggccaactc agcttttgtg gagcgagtgc ggaaacgggg cttcgaggtg
     2881 gtatatatga ccgagcccat tgacgagtac tgtgtgcagc agctcaagga atttgatggg
     2941 aagagcctgg tctcagttac caaggagggt ctggagctgc ctgaggatga ggaggagaag
     3001 aagaagatgg aagagagcaa ggcaaagttt gagaacctct gcaagctcat gaaagaaatc
     3061 ttagataaga aggttgagaa ggtgacaatc tccaatagac ttgtgtcttc accttgctgc
     3121 attgtgacca gcacctacgg ctggacagcc aatatggagc ggatcatgaa agcccaggca
     3181 cttcgggaca actccaccat gggctatatg atggccaaaa agcacctgga gatcaaccct
     3241 gaccacccca ttgtggagac gctgcggcag aaggctgagg ccgacaagaa tgataaggca
     3301 gttaaggacc tggtggtgct gctgtttgaa accgccctgc tatcttctgg cttttccctt
     3361 gaggatcccc agacccactc caaccgcatc tatcgcatga tcaagctagg tctaggtatt
     3421 gatgaagatg aagtggcagc agaggaaccc aatgctgcag ttcctgatga gatcccccct
     3481 ctcgagggcg atgaggatgc gtctcgcatg gaagaagtcg attagtcgac ggtaccgcgg
     3541 gcccgggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     3601 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3661 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3721 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3781 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3841 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3901 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3961 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     4021 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     4081 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4141 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4201 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     4261 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4321 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4381 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4441 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4501 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4561 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4621 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4681 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4741 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4801 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4861 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4921 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4981 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5041 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5101 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5161 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5221 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5281 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     5341 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5401 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5461 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5521 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5581 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5641 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5701 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     5761 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5821 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5881 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5941 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     6001 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     6061 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6121 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6181 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6241 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6301 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     6361 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6421 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6481 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6541 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6601 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6661 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6721 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6781 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6841 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
