LOCUS       sequence-223523-        7077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid expresses human HSF1 K298R mutant, which cannot undergo
            sumoylation on K298..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7077)
  AUTHORS   Hietakangas V, Ahlskog JK, Jakobsson AM, Hellesuo M, Sahlberg NM, 
            Holmberg CI, Mikhailov A, Palvimo JJ, Pirkkala L, Sistonen L
  TITLE     Phosphorylation of serine 303 is a prerequisite for the 
            stress-inducible SUMO modification of heat shock factor 1.
  JOURNAL   Mol Cell Biol. 2003 Apr . 23(8):2953-68.
  PUBMED    12665592
REFERENCE   2  (bases 1 to 7077)
  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. 2003 Apr . 23(8):2953-68."
COMMENT     Sequence Label: hHSF1 K298R
FEATURES             Location/Qualifiers
     source          1..7077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2178..2549
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             2562..2591
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2607..2624
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2668..2685)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2674..2898
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2944..3372
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3031..3050)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3241..3262
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3381..3401)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3386..3715
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3566..3701
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3628..3647
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3782..4576
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3836..3855)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4446..4465
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4750..4871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4787..4806)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4841..4860
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4920..4936)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4920..4936)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4933..4955)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4944..4960
                     /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)."
     promoter        complement(join(4968..4974,4975..4992,4993..4998))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5013..5034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5151..5168)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5322..5910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5402..5421)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6081..6872,6873..6941))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6704..6723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6942..7046)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattcg
      961 agatggatct gcccgtgggc cccggcgcgg cggggcccag caacgtcccg gccttcctga
     1021 ccaagctgtg gaccctcgtg agcgacccgg acaccgacgc gctcatctgc tggagcccga
     1081 gcgggaacag cttccacgtg ttcgaccagg gccagtttgc caaggaggtg ctgcccaagt
     1141 acttcaagca caacaacatg gccagcttcg tgcggcagct caacatgtat ggcttccgga
     1201 aagtggtcca catcgagcag ggcggcctgg tcaagccaga gagagacgac acggagttcc
     1261 agcacccatg cttcctgcgt ggccaggagc agctccttga gaacatcaag aggaaagtga
     1321 ccagtgtgtc caccctgaag agtgaagaca taaagatccg ccaggacagc gtcaccaagc
     1381 tgctgacgga cgtgcagctg atgaagggga agcaggagtg catggactcc aagctcctgg
     1441 ccatgaagca tgagaatgag gctctgtggc gggaggtggc cagccttcgg cagaagcatg
     1501 cccagcaaca gaaagtcgtc aacaagctca ttcagttcct gatctcactg gtgcagtcaa
     1561 accggatcct gggggtgaag agaaagatcc ccctgatgct gaacgacagt ggctcagcac
     1621 attccatgcc caagtatagc cggcagttct ccctggagca cgtccacggc tcgggcccct
     1681 actcggcccc ctccccagcc tacagcagct ccagcctcta cgcccctgat gctgtggcca
     1741 gctctggacc catcatctcc gacatcaccg agctggctcc tgccagcccc atggcctccc
     1801 ccggcgggag catagacgag aggcccctat ccagcagccc cctggtgcgt gtcagggagg
     1861 agccccccag cccgcctcag agcccccggg tagaggaggc gagtcccggg cgcccatctt
     1921 ccgtggacac cctcttgtcc ccgaccgccc tcattgactc catcctgcgg gagagtgaac
     1981 ctgcccccgc ctccgtcaca gccctcacgg acgccagggg ccacacggac accgagggcc
     2041 ggcctccctc ccccccgccc acctccaccc ctgaaaagtg cctcagcgta gcctgcctgg
     2101 acaagaatga gctcagtgac cacttggatg ctatggactc caacctggat aacctgcaga
     2161 ccatgctgag cagccacggc ttcagcgtgg acaccagtgc cctgctggac ctgttcagcc
     2221 cctcggtgac cgtgcccgac atgagcctgc ctgaccttga cagcagcctg gccagtatcc
     2281 aagagctcct gtctccccag gagcccccca ggcctcccga ggcagagaac agcagcccgg
     2341 attcagggaa gcagctggtg cactacacag cgcagccgct gttcctgctg gaccccggct
     2401 ccgtggacac cgggagcaac gacctgccgg tgctgtttga gctgggagag ggctcctact
     2461 tctccgaagg ggacggcttc gccgaggacc ccaccatctc cctgctgaca ggctcggagc
     2521 ctcccaaagc caaggacccc actgtctcca agcttgggcc cgaacaaaaa ctcatctcag
     2581 aagaggatct gaatagcgcc gtcgaccatc atcatcatca tcattgagtt taaacggtct
     2641 ccagcttaag tttaaaccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     2701 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2761 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2821 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     2881 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     2941 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3001 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3061 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3121 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3181 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3241 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3301 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3361 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3421 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3481 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3541 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3601 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3661 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3721 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     3781 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3841 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3901 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3961 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4021 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4081 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4141 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4201 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4261 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4321 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4381 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4441 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4501 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4561 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4621 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4681 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4741 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4801 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4861 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4921 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4981 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5041 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5101 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5161 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5221 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5281 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5341 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5401 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5461 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5521 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5581 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5641 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5701 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5761 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5821 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5881 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5941 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6001 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6061 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6121 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6181 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6241 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6301 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6361 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6421 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6481 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6541 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6601 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6661 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6721 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6781 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6841 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6901 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6961 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7021 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
