LOCUS       sequence-227544-        7077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid expresses human HSF1 S303A and S307A mutant, which 
            cannot undergo sumoylation on K298 due to disrupted PDSM motif..
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 S303/7A
FEATURES             Location/Qualifiers
     source          1..7077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1944..2315
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             2328..2357
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2373..2390
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2434..2451)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2440..2664
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2710..3138
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2797..2816)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3007..3028
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3147..3167)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3152..3481
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3332..3467
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3394..3413
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3548..4342
                     /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(3602..3621)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4212..4231
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4516..4637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4553..4572)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4607..4626
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4686..4702)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4686..4702)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4699..4721)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4710..4726
                     /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(4734..4740,4741..4758,4759..4764))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4779..4800
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4917..4934)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5088..5676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5168..5187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5847..6638,6639..6707))
                     /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     6470..6489
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6708..6812)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6887..6906)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacgggccc tctagactcg agcggccgcc actgtgctgg atatctgcag
      721 aattcgagat ggatctgccc gtgggccccg gcgcggcggg gcccagcaac gtcccggcct
      781 tcctgaccaa gctgtggacc ctcgtgagcg acccggacac cgacgcgctc atctgctgga
      841 gcccgagcgg gaacagcttc cacgtgttcg accagggcca gtttgccaag gaggtgctgc
      901 ccaagtactt caagcacaac aacatggcca gcttcgtgcg gcagctcaac atgtatggct
      961 tccggaaagt ggtccacatc gagcagggcg gcctggtcaa gccagagaga gacgacacgg
     1021 agttccagca cccatgcttc ctgcgtggcc aggagcagct ccttgagaac atcaagagga
     1081 aagtgaccag tgtgtccacc ctgaagagtg aagacataaa gatccgccag gacagcgtca
     1141 ccaagctgct gacggacgtg cagctgatga aggggaagca ggagtgcatg gactccaagc
     1201 tcctggccat gaagcatgag aatgaggctc tgtggcggga ggtggccagc cttcggcaga
     1261 agcatgccca gcaacagaaa gtcgtcaaca agctcattca gttcctgatc tcactggtgc
     1321 agtcaaaccg gatcctgggg gtgaagagaa agatccccct gatgctgaac gacagtggct
     1381 cagcacattc catgcccaag tatagccggc agttctccct ggagcacgtc cacggctcgg
     1441 gcccctactc ggccccctcc ccagcctaca gcagctccag cctctacgcc cctgatgctg
     1501 tggccagctc tggacccatc atctccgaca tcaccgagct ggctcctgcc agccccatgg
     1561 cctcccccgg cgggagcata gacgagaggc ccctatccag cagccccctg gtgcgtgtca
     1621 aggaggagcc ccccgccccg cctcaggccc cccgggtaga ggaggcgagt cccgggcgcc
     1681 catcttccgt ggacaccctc ttgtccccga ccgccctcat tgactccatc ctgcgggaga
     1741 gtgaacctgc ccccgcctcc gtcacagccc tcacggacgc caggggccac acggacaccg
     1801 agggccggcc tccctccccc ccgcccacct ccacccctga aaagtgcctc agcgtagcct
     1861 gcctggacaa gaatgagctc agtgaccact tggatgctat ggactccaac ctggataacc
     1921 tgcagaccat gctgagcagc cacggcttca gcgtggacac cagtgccctg ctggacctgt
     1981 tcagcccctc ggtgaccgtg cccgacatga gcctgcctga ccttgacagc agcctggcca
     2041 gtatccaaga gctcctgtct ccccaggagc cccccaggcc tcccgaggca gagaacagca
     2101 gcccggattc agggaagcag ctggtgcact acacagcgca gccgctgttc ctgctggacc
     2161 ccggctccgt ggacaccggg agcaacgacc tgccggtgct gtttgagctg ggagagggct
     2221 cctacttctc cgaaggggac ggcttcgccg aggaccccac catctccctg ctgacaggct
     2281 cggagcctcc caaagccaag gaccccactg tctccaagct tgggcccgaa caaaaactca
     2341 tctcagaaga ggatctgaat agcgccgtcg accatcatca tcatcatcat tgagtttaaa
     2401 cggtctccag cttaagttta aaccgctgat cagcctcgac tgtgccttct agttgccagc
     2461 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2521 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2581 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2641 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     2701 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2761 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2821 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2881 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2941 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3001 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3061 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3121 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3181 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3241 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3301 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3361 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3421 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3481 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     3541 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3601 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3661 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3721 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3781 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3841 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3901 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3961 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4021 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4081 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4141 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4201 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4261 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4321 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4381 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4441 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4501 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4561 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4621 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     4681 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4741 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4801 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4861 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4921 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4981 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5041 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5101 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5161 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5221 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5281 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5341 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5401 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5461 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5521 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5581 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5641 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5701 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5761 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5821 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5881 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5941 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6001 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6061 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6121 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6181 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6241 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6301 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6361 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6421 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6481 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6541 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6601 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6661 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6721 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6781 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6841 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     6901 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     6961 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     7021 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
