LOCUS       sequence-393126-        8301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of truncated HSF2 (DBD).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8301)
  AUTHORS   Price RM, Budzynski MA, Shen J, Mitchell JE, Kwan JZJ, Teves SS
  TITLE     Heat shock transcription factors demonstrate a distinct mode of 
            interaction with mitotic chromosomes.
  JOURNAL   Nucleic Acids Res. 2023 Apr 28:gkad304. doi: 10.1093/nar/gkad304.
  PUBMED    37114996
REFERENCE   2  (bases 1 to 8301)
  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: "Nucleic 
            Acids Res. 2023 Apr 28:gkad304. doi: 10.1093/nar/gkad304."
COMMENT     Sequence Label: Halo-H2DBD
FEATURES             Location/Qualifiers
     source          1..8301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(60..80)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    122..243
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(159..178)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     213..232
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   1005..1039
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(1530..1546)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1530..1546)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1543..1565)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1554..1570
                     /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(1578..1584,1585..1602,1603..1608))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1623..1644
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1761..1778)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1932..2520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2012..2031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2691..3482,3483..3551))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEVDKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3314..3333
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3552..3656)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3682..4137)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3819..3840)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4031..4050
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4264..4286
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4278..4295
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4279..4295
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   4425..4487
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        4940..5151
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5164..5432
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      5458..5467
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5467..5532
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5542..6432
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             6445..6465
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     misc_feature    6908..7480
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7075..7092)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7302..7321
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7489..8283
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLAATGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7543..7562)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8153..8172
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
       61 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgcgttaact
      121 aaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
      181 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
      241 ttatcatgtc tggaattgac tcaaatgatg tcaattagtc tatcagaagc tcatctggtc
      301 tcccttccgg gggacaagac atccctgttt aatatttaaa cagcagtgtt cccaaactgg
      361 gttcttatat cccttgctct ggtcaaccag gttgcagggt ttcctgtcct cacaggaacg
      421 aagtccctaa agaaacagtg gcagccaggt ttagccccgg aattgactgg attccttttt
      481 tagggcccat tggtatggct ttttccccgt atccccccag gtgtctgcag gctcaaagag
      541 cagcgagaag cgttcagagg aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc
      601 ccgcacgctg ccggctcggg gatgcggggg gagcgccgga ccggagcgga gccccgggcg
      661 gctcgctgct gccccctagc gggggaggga cgtaattaca tccctggggg ctttgggggg
      721 ggctgtccct gatatctata acaagaaaat atatatataa taagttatca cgtaagtaga
      781 acatgaaata acaatataat tatcgtatga gttaaatcat aaaagtcacg taaaagataa
      841 ccatgcgtca ttttgactca cgcggtcgtt atagttcaaa atcagtgaca cttaccgcat
      901 tgacaagcac gcctcacggg agctccaagc ggcgactgag atgtcctaaa tgcacagcga
      961 cggattcgcg ctatttagaa agagagagca atatttcaag aatgcatgcg tcaattttac
     1021 gcagactatc tttctagggt taatctagct gcatcaggat catatcgtcg ggtctttttt
     1081 ccggctcagt catcgcccaa gctggcgcta tctgggcatc ggggaggaag aagcccgtgc
     1141 ctttacccgc gaggttgaag cggcatggaa agagtttgcc gaggatgact gctgctgcat
     1201 tgacgttgag cgaaaacgca cgtttaccat gatgattcgg gaaggtgtgg ccatgcacgc
     1261 ctttaacggt gaactgttcg ttcaggccac ctgggatacc agttcgtcgc ggcttttccg
     1321 gacacagttc cggatggtca gcccgaagcg catcagcaac ccgaacaata ccggcgacag
     1381 ccggaactgc cgtgccggtg tgcagattaa tggcagcggt gcggcgctgg gatattacgt
     1441 cagcgaggac gggtatcctg gctggatgcc gcagaaatgg acatggatac cccgtgagtt
     1501 acccggcggg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     1561 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     1621 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     1681 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     1741 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     1801 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     1861 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1921 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgatgctca
     1981 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2041 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2101 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2161 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2221 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2281 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2341 aagtggtggc ctaactgcgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2401 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2461 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2521 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2581 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2641 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     2701 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     2761 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     2821 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     2881 ggaagggccg agcgcagaag tggtcctgca actttatcca cctccatcca gtctattaat
     2941 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3001 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3061 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3121 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3181 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3241 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3301 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3361 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3421 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3481 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     3541 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     3601 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     3661 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     3721 atttttgtta agtcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3781 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3841 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3901 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3961 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4021 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4081 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     4141 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     4201 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     4261 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcctcgttc attcacgttt
     4321 ttgaacccgt ggaggacggg cagactcgcg gtgcaaatgt gttttacagc gtgatggagc
     4381 agatgaagat gctcgacacg ctgcagaaca cgcagctaga ttaaccctag aaagataatc
     4441 atattgtgac gtacgttaaa gataatcatg tgtaaaattg acgcatgtgt tttatcggtc
     4501 tgtatatcga ggtttattta ttaatttgaa tagatattaa gttttattat atttacactt
     4561 acatactaat aataaattca acaaacaatt tatttatgtt tatttattta ttaaaaaaaa
     4621 caaaaactca aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc
     4681 aaagccccca gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg
     4741 gggctccgct ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag
     4801 cccgggcacg gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt
     4861 gagcctgcag acacctgggg ggatacgggg aaaaggcctc cacggccaag gatctgcgat
     4921 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
     4981 ggaggggccg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
     5041 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
     5101 gtagtcgccg tgaacgttcc ttttcgcaac gggtttgccg ccagaacaca gctgaagctt
     5161 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc
     5221 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc
     5281 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta
     5341 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt
     5401 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tactctagcc
     5461 accatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
     5521 gacgatgaca agcttggctc catggcagaa atcggtactg gctttccatt cgacccccat
     5581 tatgtggaag tcctgggcga gcgcatgcac tacgtcgatg ttggtccgcg cgatggcacc
     5641 cctgtgctgt tcctgcacgg taacccgacc tcctcctacg tgtggcgcaa catcatcccg
     5701 catgttgcac cgacccatcg ctgcattgct ccagacctga tcggtatggg caaatccgac
     5761 aaaccagacc tgggttattt cttcgacgac cacgtccgct tcatggatgc cttcatcgaa
     5821 gccctgggtc tggaagaggt cgtcctggtc attcacgact ggggctccgc tctgggtttc
     5881 cactgggcca agcgcaatcc agagcgcgtt aaaggtattg catttatgga gttcatccgc
     5941 cctatcccga cctgggacga atggccagaa tttgcccgcg agaccttcca ggccttccgc
     6001 accaccgacg tcggccgcaa gctgatcatc gatcagaacg tttttatcga gggtacgctg
     6061 ccgatgggtg tcgtccgccc gctgactgaa gtcgagatgg accattaccg cgagccgttc
     6121 ctgaatcctg ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc aatcgccggt
     6181 gagccagcga acatcgtcgc gctggtcgaa gaatacatgg actggctgca ccagtcccct
     6241 gtcccgaagc tgctgttctg gggcacccca ggcgttctga tcccaccggc cgaagccgct
     6301 cgcctggcca aaagcctgcc taactgcaag gctgtggaca tcggcccggg tctgaatctg
     6361 ctgcaagaag acaacccgga cctgatcggc agcgagatcg cgcgctggct gtcgacgctc
     6421 gagatttccg gcgagccaac cactgaggat ctgtactttc agagcgataa ctctagaatg
     6481 aagcagagtt ccaacgtgcc ggctttcctc agcaagctgt ggacgcttgt ggaggaaacc
     6541 cacaccaacg agttcatcac ctggagtcag aatggacaaa gttttctggt cttggatgag
     6601 caaagatttg caaaggaaat tcttcctaag tacttcaaac acaataacat ggcgagcttt
     6661 gtgagacaac taaatatgta tggcttccga aaagtagtgc atatcgaatc tggaattatc
     6721 aaacaggaaa gagatggccc tgttgaattt cagcatcctt atttcaagca aggccaggat
     6781 gacctgttgg agaacattaa aaggaaggtt tcatcttcaa aaccagagga aaataaaatt
     6841 cgtggtggcg gtggctcgaa acgtaaaagg cctctacttc taaacacaaa tggagcccca
     6901 aagtaagccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     6961 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     7021 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     7081 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     7141 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     7201 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     7261 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     7321 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     7381 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     7441 accacgggga cgtggttttc ctttgaaaaa cacgataata cctccggaat gattgaacaa
     7501 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     7561 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     7621 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     7681 gcgcggctat cgtggctggc cgcgacgggc gttccttgcg cagctgtgct cgacgttgtc
     7741 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     7801 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     7861 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     7921 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     7981 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     8041 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     8101 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     8161 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     8221 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     8281 tgagtcgaca atcaacctct g
//
