LOCUS       sequence-311665-        8125 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express N-ter FLAG HA tagged DHX36 isoform 2 with E335A mutation in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8125)
  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 8125)
  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-DHX36iso2E335A
FEATURES             Location/Qualifiers
     source          1..8125
                     /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(3843..3860)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3849..4073
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4206..4225)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4357..4404
                     /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             4412..5434
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4412..5434
                     /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    5564..5685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5601..5620)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5655..5674
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5734..5750)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5734..5750)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5747..5769)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5758..5774
                     /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(5782..5788,5789..5806,5807..5812))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5827..5848
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5965..5982)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6136..6724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6216..6235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6895..7686,7687..7755))
                     /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     7518..7537
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7756..7860)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7935..7954)
                     /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 ttggcggtac 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 atcagttaac cagacacagg tgtttaaaag aacccctcct ggtgttcgga
     2401 aaatagtaat tgctaccaac attgcggaga ctagcattac catagatgat gtcgtttatg
     2461 tgatagatgg aggaaaaata aaagagacgc attttgatac tcagaacaat atcagtacaa
     2521 tgtccgctga gtgggttagt aaagctaatg ccaaacagag aaaaggtcga gctggaagag
     2581 ttcaacctgg tcattgctat catctgtata atggtcttag agcaagtctt ctagatgact
     2641 atcaactgcc agaaattttg agaactcctt tggaagaact ttgtttacaa ataaagattt
     2701 taaggctagg tggaattgct tattttctga gtagattaat ggacccacca tcaaatgagg
     2761 cagtgttact ctccataaga cacctgatgg agctgaacgc tttggataaa caagaagaat
     2821 tgacgcctct tggagtccac ttggcacgat tacccgttga gccacatatt ggaaaaatga
     2881 ttctttttgg agcactgttc tgctgcttag acccagtact cactattgct gctagtctca
     2941 gtttcaaaga tccatttgtc attccactgg gaaaagaaaa gattgcagat gcaagaagaa
     3001 aggaattggc aaaggatact agaagtgatc acttaacagt tgtgaatgcg tttgagggct
     3061 gggaagaggc taggcgacgt ggtttcagat acgaaaagga ctattgctgg gaatattttc
     3121 tgtcttcaaa cacactgcag atgctgcata acatgaaagg acagtttgct gagcatcttc
     3181 ttggagctgg atttgtaagc agtagaaatc ctaaagatcc agaatctaat ataaattcag
     3241 ataatgagaa gataattaaa gctgtcatct gtgctggttt atatcccaaa gttgctaaaa
     3301 ttcgactaaa tttgggtaaa aaaagaaaaa tggtaaaagt ttacacaaaa accgatggcc
     3361 tggttgctgt tcatcctaaa tctgttaatg tggagcaaac agactttcac tacaactggc
     3421 ttatctatca cctaaagatg agaacaagca gtatatactt gtatgactgc acagaggttt
     3481 ccccatactg tctcttgttt tttggaggtg acatttccat ccagaaggat aacgatcagg
     3541 aaactattgc tgtagatgag tggattgtat ttcagtctcc agcaagaatt gcccatcttg
     3601 ttaaggaatt aagaaaggaa ctagatattc ttctgcaaga gaagattgaa agtcctcatc
     3661 ctgtagactg gaatgacact aaatccagag actgtgcagt actgtcagct attatagact
     3721 tgatcaaaac acaggaaaag gcaactccca ggaactttcc gccacgattc caggatggat
     3781 attacagctg actcgaggcg gccgcgtagg taactgaggg cccgtttaaa cccgctgatc
     3841 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3901 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3961 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4021 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     4081 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4141 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4201 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4261 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4321 caaaaaactt gattagggtg atggttcacg tacctagaag ttcctattcc gaagttccta
     4381 ttctctagaa agtataggaa cttccttggc caaaaagcct gaactcaccg cgacgtctgt
     4441 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     4501 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     4561 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc
     4621 gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat
     4681 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     4741 tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag
     4801 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     4861 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     4921 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     4981 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     5041 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     5101 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     5161 gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct
     5221 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     5281 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     5341 cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     5401 acgccccagc actcgtccga gggcaaagga atagcacgta ctacgagatt tcgattccac
     5461 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5521 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5581 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5641 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5701 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5761 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5821 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5881 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5941 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6001 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6061 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6121 cgcgttgctg gcgtttttcc ataggctccg cctccctgac gagcatcaca aaaatcgacg
     6181 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6241 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6301 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6361 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6421 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6481 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6541 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6601 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6661 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6721 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6781 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6841 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6901 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6961 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7021 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7081 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7141 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7201 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7261 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7321 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7381 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7441 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7501 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7561 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga ggtccagttc
     7621 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7681 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7741 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7801 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7861 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7921 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     7981 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     8041 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     8101 gatgtacggg ccagatatac gcgtt
//
