LOCUS       sequence-275750-        6959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral 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 6959)
  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 6959)
  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: pLenti CMV/TO Zeo dn-cHSF1
FEATURES             Location/Qualifiers
     source          1..6959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1802..1918
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1941..2244
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2245..2448
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2399..2419
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2450..2468
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2471..2489
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2530..2554
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3709..3733)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3764..4352
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3817..3837)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4235..4246)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4403..4423)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4408..4737
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4588..4723
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4749..4796
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4815..5189
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     LTR             5282..5515
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5587..5708
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5624..5643)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5678..5697
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5903..6491
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6392..6411
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6645..6662
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6779..6800
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6815..6820,6821..6838,6839..6845)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6853..6869
                     /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)."
     primer_bind     6858..6880
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6877..6893
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6877..6893
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6912..6932
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6914..6932
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa
     1801 cttttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     1861 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaattc aaaattttat
     1921 cgataagctt gggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     1981 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     2041 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     2101 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     2161 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     2221 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2281 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2341 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2401 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt
     2461 gatagagatc tccctatcag tgatagagat cgtcgactag tccagtgtgg tggaattctg
     2521 cagatatcaa caagtttgta caaaaaagca ggctttaaag gaaccaattc agtcgactgg
     2581 atccggtacc accatggatc tgcccgtggg ccccggcgcg gcggggccca gcaacgtccc
     2641 ggccttcctg accaagctgt ggaccctcgt gagcgacccg gacaccgacg cgctcatctg
     2701 ctggagcccg agcgggaaca gcttccacgt gttcgaccag ggccagtttg ccaaggaggt
     2761 gctgcccaag tacttcaagc acaacaacat ggccagcttc gtgcggcagc tcaacatgta
     2821 tggcttccgg aaagtggtcc acatcgagca gggcggcctg gtcaagccag agagagacga
     2881 cacggagttc cagcacccat gcttcctgcg tggccaggag cagctccttg agaacatcaa
     2941 gaggaaagtg accagtgtgt ccaccctgaa gagtgaagac ataaagatcc gccaggacag
     3001 cgtcaccaag ctgctgacgg acgtgcagct gatgaagggg aagcaggagt gcatggactc
     3061 caagctcctg gccatgaagc atgagaatga ggctctgtgg cgggaggtgg ccagccttcg
     3121 gcagaagcat gcccagcaac agaaagtcat cctgggggtg aagagaaaga tccccctgat
     3181 gctgaacgac agtggctcag cacattccat gcccaagtat agccggcagt tctccctgga
     3241 gcacgtccac ggctcgggcc cctactcggc cccctcccca gcctacagca gctccagcct
     3301 ctacgcccct gatgctgtgg ccagctctgg acccatcatc tccgacatca ccgagctggc
     3361 tcctgccagc cccatggcct cccccggcgg gagcatagac gagaggcccc tatccagcag
     3421 ccccctggtg cgtgtcaagg aggagccccc cagcccgcct cagagccccc gggtagagga
     3481 ggcgagtccc gggcgcccat cttccgtgga caccctcttg tccccgaccg ccctcattga
     3541 ctccatcctg cgggagagtg aacctgcccc cgcctccgtc acagccctca cggacgccag
     3601 gggccacacg gacaccgagg gccggcctcc ctcccccccg cccacctcca cccctgaaaa
     3661 gtgcctcagc gtagcctgcc tgtaggcggc cgcactcgag atatctagac ccagctttct
     3721 tgtacaaagt ggttgatatc cagcacagtg gcggccgctc gacaatcaac ctctggatta
     3781 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3841 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3901 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3961 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     4021 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     4081 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     4141 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     4201 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     4261 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     4321 gagtcggatc tccctttggg ccgcctcccc gcctggaatt ctgcagatat ccggttagta
     4381 atgagtttgg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4441 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4501 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4561 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4621 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4681 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4741 ctccccctgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     4801 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     4861 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     4921 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     4981 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     5041 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     5101 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     5161 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttaaat ggtaccttta
     5221 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     5281 ctggaagggc tagctcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     5341 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     5401 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     5461 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     5521 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     5581 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5641 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5701 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa
     5761 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5821 aggccgaggc cgccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5881 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5941 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6001 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6061 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6121 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6181 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6241 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6301 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6361 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6421 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6481 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6541 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6601 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6661 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     6721 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     6781 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     6841 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     6901 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaa
//
