LOCUS       sequence-258256-        6324 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 6324)
  AUTHORS   Wang X, Grammatikakis N, Siganou A, Calderwood SK
  TITLE     Regulation of molecular chaperone gene transcription involves the 
            serine phosphorylation, 14-3-3 epsilon binding, and cytoplasmic 
            sequestration of heat shock factor 1.
  JOURNAL   Mol Cell Biol. 2003 Sep;23(17):6013-26.
  PUBMED    12917326
REFERENCE   2  (bases 1 to 6324)
  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: "Mol Cell 
            Biol."; date: "2003-09"; volume: "23(17)"; pages: "6013-26"
COMMENT     Sequence Label: HSF1-GFPN3
FEATURES             Location/Qualifiers
     source          1..6324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1790..2161
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     regulatory      2204..2213
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2210..2212,2213..2215,2216..2929)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2210..2212,2213..2215,2216..2926)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2255..2276)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2516..2535)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2863..2884
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3052..3173
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3089..3108)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3143..3162
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3180..3635)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3317..3338)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3529..3548
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3662..3766
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3768..4125
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3768..3959
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3791..3811)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3976..4111
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4160..4954
                     /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             4160..4954
                     /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(4214..4233)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4824..4843
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5142..5161)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5186..5233
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5562..6150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6051..6070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttggg taccatggat ctgcccgtgg gccccggcgc
      601 ggcggggccc agcaacgtcc cggccttcct gaccaagctg tggaccctcg tgagcgaccc
      661 ggacaccgac gcgctcatct gctggagccc gagcgggaac agcttccacg tgttcgacca
      721 gggccagttt gccaaggagg tgctgcccaa gtacttcaag cacaacaaca tggccagctt
      781 cgtgcggcag ctcaacatgt atggcttccg gaaagtggtc cacatcgagc agggcggcct
      841 ggtcaagcca gagagagacg acacggagtt ccagcaccca tgcttcctgc gtggccagga
      901 gcagctcctt gagaacatca agaggaaagt gaccagtgtg tccaccctga agagtgaaga
      961 cataaagatc cgccaggaca gcgtcaccaa gctgctgacg gacgtgcagc tgatgaaggg
     1021 gaagcaggag tgcatggact ccaagctcct ggccatgaag catgagaatg aggctctgtg
     1081 gcgggaggtg gccagccttc ggcagaagca tgcccagcaa cagaaagtcg tcaacaagct
     1141 cattcagttc ctgatctcac tggtgcagtc aaaccggatc ctgggggtga agagaaagat
     1201 ccccctgatg ctgaacgaca gtggctcagc acattccatg cccaagtata gccggcagtt
     1261 ctccctggag cacgtccacg gctcgggccc ctactcggcc ccctccccag cctacagcag
     1321 ctccagcctc tacgcccctg atgctgtggc cagctctgga cccatcatct ccgacatcac
     1381 cgagctggct cctgccagcc ccatggcctc ccccggcggg agcatagacg agaggcccct
     1441 atccagcagc cccctggtgc gtgtcaagga ggagcccccc agcccgcctc agagcccccg
     1501 ggtagaggag gcgagtcccg ggcgcccatc ttccgtggac accctcttgt ccccgaccgc
     1561 cctcattgac tccatcctgc gggagagtga acctgccccc gcctccgtca cagccctcac
     1621 ggacgccagg ggccacacgg acaccgaggg ccggcctccc tcccccccgc ccacctccac
     1681 ccctgaaaag tgcctcagcg tagcctgcct ggacaagaat gagctcagtg accacttgga
     1741 tgctatggac tccaacctgg ataacctgca gaccatgctg agcagccacg gcttcagcgt
     1801 ggacaccagt gccctgctgg acctgttcag cccctcggtg accgtgcccg acatgagcct
     1861 gcctgacctt gacagcagcc tggccagtat ccaagagctc ctgtctcccc aggagccccc
     1921 caggcctccc gaggcagaga acagcagccc ggattcaggg aagcagctgg tgcactacac
     1981 agcgcagccg ctgttcctgc tggaccccgg ctccgtggac accgggagca acgacctgcc
     2041 ggtgctgttt gagctgggag agggctccta cttctccgaa ggggacggct tcgccgagga
     2101 ccccaccatc tccctgctga caggctcgga gcctcccaaa gccaaggacc ccactgtctc
     2161 cgagaattct gcagtcgacg gtaccgcggg cccgggatcc atcgccacca tggtgagcaa
     2221 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2281 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     2341 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2401 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     2461 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2521 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2581 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2641 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     2701 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     2761 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     2821 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     2881 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga
     2941 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     3001 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3061 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3121 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     3181 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3241 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3301 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3361 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3421 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3481 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3541 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3601 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3661 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3721 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3781 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3841 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3901 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3961 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4021 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4081 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     4141 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4201 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4261 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4321 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4381 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4441 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4501 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4561 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4621 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4681 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4741 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4801 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4861 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4921 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4981 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5041 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5101 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     5161 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     5221 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5281 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5341 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     5401 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5461 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5521 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5581 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5641 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5701 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5761 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5821 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5881 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5941 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6001 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6061 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6121 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6181 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6241 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
     6301 tcatagccca tatatggagt tccg
//
