LOCUS       sequence-271440-        3871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for constitutively active dominant-negative 
            HSF1 fused to the DHFR destabilized domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3871)
  AUTHORS   Moore CL, Dewal MB, Nekongo EE, Santiago S, Lu NB, Levine SS, 
            Shoulders MD
  TITLE     Transportable, Chemical Genetic Methodology for the Small 
            Molecule-Mediated Inhibition of Heat Shock Factor 1.
  JOURNAL   ACS Chem Biol. 2016 Jan 15;11(1):200-10. doi: 
            10.1021/acschembio.5b00740. Epub 2015 Nov 19.
  PUBMED    26502114
REFERENCE   2  (bases 1 to 3871)
  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"; doi: 
            "10.1021/acschembio.5b00740"; journalName: "ACS Chem Biol"; date: 
            "2016-01-15- 15"; volume: "11"; issue: "1"; pages: "200-10"
COMMENT     Sequence Label: pENTR1A DHFR.dn-cHSF1
FEATURES             Location/Qualifiers
     source          1..3871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    67..166
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1306..1782)
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="MISLIAALAVDYVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH
                     TWESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVIEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     protein_bind    complement(1809..1908)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1927..1950)
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     terminator      complement(1958..1985)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2077..2163)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2328..2916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2408..2427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3006..3815)
                     /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     3729..3748
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3807..3826
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tatttttatc ttgtgcaatg taacatcaga gattttgaga cacgggccag agctgcagct
       61 ggatggcaaa taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat
      121 aagcaatgct ttcttataat gccaactttg tacaagaaag ctgggtctag atatctcgag
      181 tgcggccgcc tacaggcagg ctacgctgag gcacttttca ggggtggagg tgggcggggg
      241 ggagggaggc cggccctcgg tgtccgtgtg gcccctggcg tccgtgaggg ctgtgacgga
      301 ggcgggggca ggttcactct cccgcaggat ggagtcaatg agggcggtcg gggacaagag
      361 ggtgtccacg gaagatgggc gcccgggact cgcctcctct acccgggggc tctgaggcgg
      421 gctggggggc tcctccttga cacgcaccag ggggctgctg gataggggcc tctcgtctat
      481 gctcccgccg ggggaggcca tggggctggc aggagccagc tcggtgatgt cggagatgat
      541 gggtccagag ctggccacag catcaggggc gtagaggctg gagctgctgt aggctgggga
      601 gggggccgag taggggcccg agccgtggac gtgctccagg gagaactgcc ggctatactt
      661 gggcatggaa tgtgctgagc cactgtcgtt cagcatcagg gggatctttc tcttcacccc
      721 caggatgact ttctgttgct gggcatgctt ctgccgaagg ctggccacct cccgccacag
      781 agcctcattc tcatgcttca tggccaggag cttggagtcc atgcactcct gcttcccctt
      841 catcagctgc acgtccgtca gcagcttggt gacgctgtcc tggcggatct ttatgtcttc
      901 actcttcagg gtggacacac tggtcacttt cctcttgatg ttctcaagga gctgctcctg
      961 gccacgcagg aagcatgggt gctggaactc cgtgtcgtct ctctctggct tgaccaggcc
     1021 gccctgctcg atgtggacca ctttccggaa gccatacatg ttgagctgcc gcacgaagct
     1081 ggccatgttg ttgtgcttga agtacttggg cagcacctcc ttggcaaact ggccctggtc
     1141 gaacacgtgg aagctgttcc cgctcgggct ccagcagatg agcgcgtcgg tgtccgggtc
     1201 gctcacgagg gtccacagct tggtcaggaa ggccgggacg ttgctgggcc ccgccgcgcc
     1261 ggggcccacg ggcagatcca tggcggatcc acttccactt cctgctcgcc gctccagaat
     1321 ctcaaagcaa tagctgtgag agttctgcgc atcagcatcg tggaattcgc tgaataccga
     1381 ttcccagtca tccggctcgt aatccgggaa atgggtgtcg ccttccactt ctgcgtcgat
     1441 atgcgtcaga tacagttttt gcgcttttgg caagaactgt tcaataacgc gaccgccgcc
     1501 aatcaccatg atttctggta cgtcaccaca cgccgcgatg gcttcatcca ccgacttcac
     1561 ccacgttacg cgatcgtccg tactcggttg actgctgagg ataatatttt tgcgtcctgg
     1621 caacggacga ccgattgatt cccaggtatg gcggcccata atcacgggtt tatttaaggt
     1681 gttgcgttta aaccaggcga gatcggcagg caggttccac ggcatggcgt tttccatgcc
     1741 gataacgtaa tctaccgcta acgccgcaat cagactgatc atggtgtcga ctgaattggt
     1801 tcctttaaag cctgcttttt tgtacaaagt tggcattata aaaaagcatt gcttatcaat
     1861 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttggg gcccgagctt
     1921 aagtaactaa ctaacaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc
     1981 cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg
     2041 ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt
     2101 caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat
     2161 ttgatgcctg gcagttccct actctcgctt agtagttaga cgtccccgag atccatgcta
     2221 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2281 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2341 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2401 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2461 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2521 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2581 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2641 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2701 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2761 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2821 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2881 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2941 gggtctgacg ctcagtggaa cggggcccaa tctgaataat gttacaacca attaaccaat
     3001 tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     3061 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     3121 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     3181 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     3241 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     3301 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     3361 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     3421 atcgaatgca accggcgcag ggacactgcc agcgcatcaa caatattttc acctgaatca
     3481 ggatattctt ctaatacctg gaatgctgtt tttccgggga tcgcagtggt gagtaaccat
     3541 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     3601 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     3661 agaaacaact ctggcgcatc gggcttccca tacaagcgat agattgtcgc acctgattgc
     3721 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     3781 cgcggcctcg acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg
     3841 taagcagaca gttttattgt tcatgatgat a
//
