LOCUS       sequence-271742-        7040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for inducible expression of a 
            constitutively active dominant-negative HSF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7040)
  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 7040)
  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: pT-Rex Dest30 dn-cHSF1
FEATURES             Location/Qualifiers
     source          1..7040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..312
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        313..516
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     467..487
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    518..536
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    539..557
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     567..591
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    699..723
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1878..1902)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1963..1982)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1964..1982)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1993..2010)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1993..2009)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2002..2024)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    2414..2548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2451..2470)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2505..2524
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2674..3129
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2761..2780)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2971..2992
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3246..3603
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3246..3437
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3269..3289)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3454..3589
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3654..4448
                     /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(3708..3727)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4318..4337
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4512..4560
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4601..4620)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4720..4742
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4866..4970
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4971..5039,5040..5831)
                     /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     complement(5189..5208)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6002..6590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6491..6510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6744..6761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(6835..6939)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atgcatgtcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
       61 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      121 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      181 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      241 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      301 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      361 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      421 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      481 aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga tagagatctc
      541 cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat cgcctggaga
      601 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccggact
      661 ctagaggatc cctaccggtg atatcctcga gcccatcaac aagtttgtac aaaaaagcag
      721 gctttaaagg aaccaattca gtcgactgga tccggtacca ccatggatct gcccgtgggc
      781 cccggcgcgg cggggcccag caacgtcccg gccttcctga ccaagctgtg gaccctcgtg
      841 agcgacccgg acaccgacgc gctcatctgc tggagcccga gcgggaacag cttccacgtg
      901 ttcgaccagg gccagtttgc caaggaggtg ctgcccaagt acttcaagca caacaacatg
      961 gccagcttcg tgcggcagct caacatgtat ggcttccgga aagtggtcca catcgagcag
     1021 ggcggcctgg tcaagccaga gagagacgac acggagttcc agcacccatg cttcctgcgt
     1081 ggccaggagc agctccttga gaacatcaag aggaaagtga ccagtgtgtc caccctgaag
     1141 agtgaagaca taaagatccg ccaggacagc gtcaccaagc tgctgacgga cgtgcagctg
     1201 atgaagggga agcaggagtg catggactcc aagctcctgg ccatgaagca tgagaatgag
     1261 gctctgtggc gggaggtggc cagccttcgg cagaagcatg cccagcaaca gaaagtcatc
     1321 ctgggggtga agagaaagat ccccctgatg ctgaacgaca gtggctcagc acattccatg
     1381 cccaagtata gccggcagtt ctccctggag cacgtccacg gctcgggccc ctactcggcc
     1441 ccctccccag cctacagcag ctccagcctc tacgcccctg atgctgtggc cagctctgga
     1501 cccatcatct ccgacatcac cgagctggct cctgccagcc ccatggcctc ccccggcggg
     1561 agcatagacg agaggcccct atccagcagc cccctggtgc gtgtcaagga ggagcccccc
     1621 agcccgcctc agagcccccg ggtagaggag gcgagtcccg ggcgcccatc ttccgtggac
     1681 accctcttgt ccccgaccgc cctcattgac tccatcctgc gggagagtga acctgccccc
     1741 gcctccgtca cagccctcac ggacgccagg ggccacacgg acaccgaggg ccggcctccc
     1801 tcccccccgc ccacctccac ccctgaaaag tgcctcagcg tagcctgcct gtaggcggcc
     1861 gcactcgaga tatctagacc cagctttctt gtacaaagtg gttgatgggc ggccgctcta
     1921 gagggcccaa gcttacgcgt gcatgcgacg tcatagctct ctccctatag tgagtcgtat
     1981 tataagctag gcactggccg tcgttttaca acgtcgtgac tgggaaaact gctagcttgg
     2041 gatctttgtg aaggaacctt acttctgtgg tgtgacataa ttggacaaac tacctacaga
     2101 gatttaaagc tctaaggtaa atataaaatt tttaagtgta taatgtgtta aactagctgc
     2161 atatgcttgc tgcttgagag ttttgcttac tgagtatgat ttatgaaaat attatacaca
     2221 ggagctagtg attctaattg tttgtgtatt ttagattcac agtcccaagg ctcatttcag
     2281 gcccctcagt cctcacagtc tgttcatgat cataatcagc cataccacat ttgtagaggt
     2341 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     2401 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2461 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2521 catcaatgta tcttatcatg tctggatcga tcctgcatta atgaatcggc caacgcgcgg
     2581 ggagaggcgg tttgcgtatt ggctggcgta atagcgaaga ggcccgcacc gatcgccctt
     2641 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     2701 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2761 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2821 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2881 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2941 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3001 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3061 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     3121 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3181 acgcggatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt
     3241 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3301 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3361 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3421 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3481 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3541 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3601 aaaaagcttg attcttctga cacaacagtc tcgaacttaa ggctagagcc accatgattg
     3661 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3721 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3781 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3841 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3901 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3961 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4021 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4081 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4141 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4201 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4261 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4321 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4381 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4441 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4501 acgatggccg caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg
     4561 aatcgatagc gataaggatc cgcgtatggt gcactctcag tacaatctgc tctgatgccg
     4621 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4681 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4741 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4801 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4861 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4921 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4981 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5041 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5101 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5161 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5221 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5281 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5341 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5401 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5461 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5521 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5581 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5641 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5701 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5761 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5821 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5881 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5941 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6001 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6061 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6121 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6661 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6721 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     6781 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagagct tgcaattcgc
     6841 gctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6901 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6961 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtattac
     7021 gaggcccttt cacgcattag
//
