LOCUS       sequence-271749-        4321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for constitutively active HSF1 fused to the 
            DHFR destabilized domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4321)
  AUTHORS   Phillips AM, Gonzalez LO, Nekongo EE, Ponomarenko AI, McHugh SM, 
            Butty VL, Levine SS, Lin YS, Mirny LA, Shoulders MD
  TITLE     Host proteostasis modulates influenza evolution.
  JOURNAL   Elife. 2017 Sep 26;6. doi: 10.7554/eLife.28652.
  PUBMED    28949290
REFERENCE   2  (bases 1 to 4321)
  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: "Elife."; 
            date: "2017-09-26"; volume: "6. doi"; pages: " 10.7554/eLife.28652"
COMMENT     Sequence Label: pENTR1A DHFR.cHSF1
FEATURES             Location/Qualifiers
     source          1..4321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             484..960
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="MISLIAALAVDYVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH
                     TWESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVIEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             2152..2523
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     protein_bind    complement(2550..2649)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2761..2780)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2772..3581
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2839..2858)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3671..4259
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4160..4179
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac
      481 gccatgatca gtctgattgc ggcgttagcg gtagattacg ttatcggcat ggaaaacgcc
      541 atgccgtgga acctgcctgc cgatctcgcc tggtttaaac gcaacacctt aaataaaccc
      601 gtgattatgg gccgccatac ctgggaatca atcggtcgtc cgttgccagg acgcaaaaat
      661 attatcctca gcagtcaacc gagtacggac gatcgcgtaa cgtgggtgaa gtcggtggat
      721 gaagccatcg cggcgtgtgg tgacgtacca gaaatcatgg tgattggcgg cggtcgcgtt
      781 attgaacagt tcttgccaaa agcgcaaaaa ctgtatctga cgcatatcga cgcagaagtg
      841 gaaggcgaca cccatttccc ggattacgag ccggatgact gggaatcggt attcagcgaa
      901 ttccacgatg ctgatgcgca gaactctcac agctattgct ttgagattct ggagcggcga
      961 gcaggaagtg gaagtggatc cgccatggat ctgcccgtgg gccccggcgc ggcggggccc
     1021 agcaacgtcc cggccttcct gaccaagctg tggaccctcg tgagcgaccc ggacaccgac
     1081 gcgctcatct gctggagccc gagcgggaac agcttccacg tgttcgacca gggccagttt
     1141 gccaaggagg tgctgcccaa gtacttcaag cacaacaaca tggccagctt cgtgcggcag
     1201 ctcaacatgt atggcttccg gaaagtggtc cacatcgagc agggcggcct ggtcaagcca
     1261 gagagagacg acacggagtt ccagcaccca tgcttcctgc gtggccagga gcagctcctt
     1321 gagaacatca agaggaaagt gaccagtgtg tccaccctga agagtgaaga cataaagatc
     1381 cgccaggaca gcgtcaccaa gctgctgacg gacgtgcagc tgatgaaggg gaagcaggag
     1441 tgcatggact ccaagctcct ggccatgaag catgagaatg aggctctgtg gcgggaggtg
     1501 gccagccttc ggcagaagca tgcccagcaa cagaaagtca tcctgggggt gaagagaaag
     1561 atccccctga tgctgaacga cagtggctca gcacattcca tgcccaagta tagccggcag
     1621 ttctccctgg agcacgtcca cggctcgggc ccctactcgg ccccctcccc agcctacagc
     1681 agctccagcc tctacgcccc tgatgctgtg gccagctctg gacccatcat ctccgacatc
     1741 accgagctgg ctcctgccag ccccatggcc tcccccggcg ggagcataga cgagaggccc
     1801 ctatccagca gccccctggt gcgtgtcaag gaggagcccc ccagcccgcc tcagagcccc
     1861 cgggtagagg aggcgagtcc cgggcgccca tcttccgtgg acaccctctt gtccccgacc
     1921 gccctcattg actccatcct gcgggagagt gaacctgccc ccgcctccgt cacagccctc
     1981 acggacgcca ggggccacac ggacaccgag ggccggcctc cctccccccc gcccacctcc
     2041 acccctgaaa agtgcctcag cgtagcctgc ctggacaaga atgagctcag tgaccacttg
     2101 gatgctatgg actccaacct ggataacctg cagaccatgc tgagcagcca cggcttcagc
     2161 gtggacacca gtgccctgct ggacctgttc agcccctcgg tgaccgtgcc cgacatgagc
     2221 ctgcctgacc ttgacagcag cctggccagt atccaagagc tcctgtctcc ccaggagccc
     2281 cccaggcctc ccgaggcaga gaacagcagc ccggattcag ggaagcagct ggtgcactac
     2341 acagcgcagc cgctgttcct gctggacccc ggctccgtgg acaccgggag caacgacctg
     2401 ccggtgctgt ttgagctggg agagggctcc tacttctccg aaggggacgg cttcgccgag
     2461 gaccccacca tctccctgct gacaggctcg gagcctccca aagccaagga ccccactgtc
     2521 tcctaggcgg ccgcactcga gatatctaga cccagctttc ttgtacaaag ttggcattat
     2581 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     2641 cattatttgc catccagctg cagctctggc ccgtgtctca aaatctctga tgttacattg
     2701 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     2761 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
     2821 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     2881 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     2941 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     3001 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     3061 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     3121 atattgttga tgcgctggca gtgtccctgc gccggttgca ttcgattcct gtttgtaatt
     3181 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     3241 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     3301 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     3361 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     3421 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     3481 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     3541 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     3601 tgtaacatta ttcagattgg gccccgttcc actgagcgtc agaccccgta gaaaagatca
     3661 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3721 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3781 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3841 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3901 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3961 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4021 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4081 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4141 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4201 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4261 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4321 t
//
