LOCUS       sequence-311664-        8167 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express N-ter FLAG HA tagged DHX36 isoform 1 with E335A mutation in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8167)
  AUTHORS   Sauer M, Juranek SA, Marks J, De Magis A, Kazemier HG, Hilbig D, 
            Benhalevy D, Wang X, Hafner M, Paeschke K
  TITLE     DHX36 prevents the accumulation of translationally inactive mRNAs 
            with G4-structures in untranslated regions.
  JOURNAL   Nat Commun. 2019 Jun 3;10(1):2421. doi: 10.1038/s41467-019-10432-5.
  PUBMED    31160600
REFERENCE   2  (bases 1 to 8167)
  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: "Nat 
            Commun."; date: "2019-06-3"; volume: "10(1):2421. doi"; pages: " 
            10.1038/s41467-019-10432-5"
COMMENT     Sequence Label: pFRT-DHX36iso1E335A
FEATURES             Location/Qualifiers
     source          1..8167
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    586..604
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    607..625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     635..659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             750..773
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             774..800
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3885..3902)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3891..4115
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4248..4267)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4399..4446
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4454..5476
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4454..5476
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5606..5727
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5643..5662)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5697..5716
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5776..5792)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5776..5792)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5789..5811)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5800..5816
                     /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(5824..5830,5831..5848,5849..5854))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5869..5890
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6007..6024)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6178..6766)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6258..6277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6937..7728,7729..7797))
                     /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     7560..7579
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7798..7902)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7977..7996)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
      601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccggactct agcgtttaaa cttaagatgg actacaagga cgacgatgac aagtacccat
      781 acgatgttcc agattacgct ggatccatga gttatgacta ccatcagaac tggggccgtg
      841 atgggggtcc ccgcagctcc ggtgggggct atggaggggg gccagcaggg ggtcatggag
      901 gtaaccgagg ctccggagga ggcggcggcg gcggaggggg tggtcgaggc ggcaggggcc
      961 ggcatcccgg gcacctgaaa ggccgcgaaa tcggcatgtg gtacgcgaaa aaacaggggc
     1021 agaagaacaa ggaagcggag aggcaagaga gagctgtagt acacatggat gaacgacgag
     1081 aagaacaaat tgtacagtta ctgaattctg ttcaagcgaa gaatgataaa gagtcagaag
     1141 cacagatatc ctggtttgct cctgaggatc atggatacgg tactgaagtt tctactaaga
     1201 acacaccatg ctcagagaac aaacttgaca tccaggaaaa gaagttgata aatcaagaaa
     1261 aaaaaatgtt tagaatcagg aacagatcat atattgaccg agattctgag tatctcttgc
     1321 aagaaaatga accagatgga actttagacc aaaaattatt ggaagattta caaaagaaaa
     1381 aaaatgacct tcggtatatt gaaatgcagc atttcagaga aaagctgcct tcgtatggaa
     1441 tgcaaaagga attggtaaat ttaattgata accatcaggt aacagtaata agtggtgaaa
     1501 ctggttgtgg caaaaccact caagttactc agttcatttt ggataactac attgaaagag
     1561 gaaaaggatc tgcttgcaga atagtttgta ctcagccaag aagaattagt gccatttcag
     1621 ttgcggaaag agtagctgca gaaagggcag aatcttgtgg cagtggtaat agtactggat
     1681 atcaaattcg tctccagagt cggttgccaa ggaaacaggg ttctatctta tactgtacaa
     1741 caggaatcat ccttcagtgg ctccagtcag acccgtattt gtccagtgtt agtcatatcg
     1801 tacttgatgc aatccatgaa agaaatctgc agtcagatgt tttaatgact gttgttaaag
     1861 accttctcaa ttttcgatct gacttgaaag taatattgat gagtgcaaca ttgaatgcag
     1921 aaaagttttc agaatatttt ggtaactgtc caatgataca tatacctggt tttacctttc
     1981 cggttgtgga atatcttttg gaagatgtaa ttgaaaaaat aaggtatgtt ccagaacaaa
     2041 aagaacacag atcccagttt aagaggggtt tcatgcaagg gcatgtaaat agacaagaaa
     2101 aagaagaaaa agaagcaata tataaagaac gttggccaga ttatgtaagg gaactgcgaa
     2161 gaaggtattc tgcaagtact gtagatgtta tagaaatgat ggaggatgat aaagttgatc
     2221 tgaatttgat tgttgccctc atccgataca ttgttttgga agaagaggat ggtgcgatac
     2281 tggtctttct gccaggctgg gacaatatca gcactttaca tgatctcttg atgtcacaag
     2341 taatgtttaa atcagataaa tttttaatta tacctttaca ttcactgatg cctacagtta
     2401 accagacaca ggtgtttaaa agaacccctc ctggtgttcg gaaaatagta attgctacca
     2461 acattgcgga gactagcatt accatagatg atgtcgttta tgtgatagat ggaggaaaaa
     2521 taaaagagac gcattttgat actcagaaca atatcagtac aatgtccgct gagtgggtta
     2581 gtaaagctaa tgccaaacag agaaaaggtc gagctggaag agttcaacct ggtcattgct
     2641 atcatctgta taatggtctt agagcaagtc ttctagatga ctatcaactg ccagaaattt
     2701 tgagaactcc tttggaagaa ctttgtttac aaataaagat tttaaggcta ggtggaattg
     2761 cttattttct gagtagatta atggacccac catcaaatga ggcagtgtta ctctccataa
     2821 gacacctgat ggagctgaac gctttggata aacaagaaga attgacacct cttggagtcc
     2881 acttggcacg attacccgtt gagccacata ttggaaaaat gattcttttt ggagcactgt
     2941 tctgctgctt agacccagta ctcactattg ctgctagtct cagtttcaaa gatccatttg
     3001 tcattccact gggaaaagaa aagattgcag atgcaagaag aaaggaattg gcaaaggata
     3061 ctagaagtga tcacttaaca gttgtgaatg cgtttgaggg ctgggaagag gctaggcgac
     3121 gtggtttcag atacgaaaag gactattgct gggaatattt tctgtcttca aacacactgc
     3181 agatgctgca taacatgaaa ggacagtttg ctgagcatct tcttggagct ggatttgtaa
     3241 gcagtagaaa tcctaaagat ccagaatcta atataaattc agataatgag aagataatta
     3301 aagctgtcat ctgtgctggt ttatatccca aagttgctaa aattcgacta aatttgggta
     3361 aaaaaagaaa aatggtaaaa gtttacacaa aaaccgatgg cctggttgct gttcatccta
     3421 aatctgttaa tgtggagcaa acagactttc actacaactg gcttatctat cacctaaaga
     3481 tgagaacaag cagtatatac ttgtatgact gcacagaggt ttccccatac tgtctcttgt
     3541 tttttggagg tgacatttcc atccagaagg ataacgatca ggaaactatt gctgtagatg
     3601 agtggattgt atttcagtct ccagcaagaa ttgcccatct tgttaaggaa ttaagaaagg
     3661 aactagatat tcttctgcaa gagaagattg aaagtcctca tcctgtagac tggaatgaca
     3721 ctaaatccag agactgtgca gtactgtcag ctattataga cttgatcaaa acacaggaaa
     3781 aggcaactcc caggaacttt ccgccacgat tccaggatgg atattacagc tgactcgagg
     3841 cggccgcgta ggtaactgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     3901 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3961 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     4021 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     4081 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     4141 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4201 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4261 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4321 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4381 tgatggttca cgtacctaga agttcctatt ccgaagttcc tattctctag aaagtatagg
     4441 aacttccttg gccaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     4501 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     4561 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     4621 ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt
     4681 gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     4741 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     4801 ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg
     4861 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     4921 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     4981 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     5041 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     5101 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     5161 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     5221 aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     5281 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     5341 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg
     5401 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     5461 gagggcaaag gaatagcacg tactacgaga tttcgattcc accgccgcct tctatgaaag
     5521 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5581 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5641 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5701 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5761 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5821 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5881 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5941 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6001 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6061 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6121 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6181 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6241 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6301 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6361 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6421 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6481 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6541 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6601 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6661 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6721 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6781 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6841 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6901 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6961 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7021 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7081 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7141 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7201 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7261 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7321 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7381 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7441 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7501 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7561 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7621 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7681 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7741 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7801 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7861 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7921 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     7981 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     8041 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     8101 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     8161 acgcgtt
//
