LOCUS       sequence-329859-        7112 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of N. furzeri DDX5-Delta-IDR(1-483) mutant in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7112)
  AUTHORS   Harel I, Chen YR, Ziv I, Singh PP, Heinzer D, Navarro Negredo P, 
            Goshtchevsky U, Wang W, Astre G, Moses E, McKay A, Machado BE, 
            Hebestreit K, Yin S, Sanchez Alvarado A, Jarosz DF, Brunet A
  TITLE     Identification of protein aggregates in the aging vertebrate brain 
            with prion-like and phase-separation properties.
  JOURNAL   Cell Rep. 2024 Jun 25;43(6):112787. doi: 
            10.1016/j.celrep.2023.112787. Epub 2024 May 28.
  PUBMED    38810650
REFERENCE   2  (bases 1 to 7112)
  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: "Cell Rep.";
            date: "2024-06-25"; volume: "43(6):112787. doi"; pages: " 
            10.1016/j.celrep.2023.112787. Epub 2024 May 28"
COMMENT     Sequence Label: pDEST17-His10-Smt3-DDX5-Delta-IDR(kf)-SNAP
FEATURES             Location/Qualifiers
     source          1..7112
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             113..142
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             155..166
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             179..472
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     protein_bind    485..509
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1977..2001)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2003..2026
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2045..2590
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for bacterial expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     primer_bind     complement(2658..2676)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2672..2719
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3085..3090,3091..3107,3108..3113))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3140..3158)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3226..3330
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3331..3399,3400..4191)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3549..3568)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4362..4950
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4851..4870
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5104..5121
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5136..5275
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5201..5220)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5361..5383
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5377..5568)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     7030..7049
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagaaataat tttgtttaac tttaagaagg agatatacat atgtcgtact accatcacca
      121 tcaccatcac catcatcatc acagcagcgg ccatatcgaa ggtcgtcata tggctagcat
      181 gtcggactca gaagtcaatc aagaagctaa gccagaggtc aagccagaag tcaagcctga
      241 gactcacatc aatttaaagg tgtccgatgg atcttcagag atcttcttca agatcaaaaa
      301 gaccactcct ttaagaaggc tgatggaagc gttcgctaaa agacagggta aggaaatgga
      361 ctccttaaga ttcttgtacg acggtattag aattcaagct gatcagaccc ctgaagattt
      421 ggacatggag gataacgata ttattgaggc tcacagagaa cagattggtg gatccctcga
      481 atcaacaagt ttgtacaaaa aagcaggctt cgaaggagat aacaaaatgc ctggatttgc
      541 agacagagac cgcggcagag atagagggta tgggggtggc ccccctcggt ttggtggcgg
      601 tgggggagga ggcggcaaca gaggaggacc tctgctggga aagtttggaa accctggtga
      661 gaggctgcgc aagaagcact ggaaccttga tgaactacca aaatttcaaa agaatttcta
      721 ccaggaacac ccagatgtca cacacagacc actgcaggag gttgaccagt accgcagaaa
      781 caaagaagtt acagttaaag gcagagattg tcccaaaccg attgtaaaat tccatgaagc
      841 tgcttttccc aactacgtca tggatgtcat tgtcaaacag aactggactg aaccaactcc
      901 gatccagtct caggggtggc ccattgtcct gagtggaaaa gacatggtcg gcatcgctca
      961 aacggggtct gggaagactt tggcatatct tctgcctgca gtcgttcaca tccaacatca
     1021 gcccttcttg gagcacggag acgggcctat ttgcttggtg ctggcaccaa cccgtgagct
     1081 ggctcagcag gtgcaacagg tggctgctga gtacggcagg gcctcccgtc tcaagagtac
     1141 atgcatctat ggtggtgctc ccaaaggacc acaaatccgg gaccttgaga ggggtgttga
     1201 gatttgcatc gccaccccag gtcgcctcat tgatttcctg gaatgtggta aaactaattt
     1261 gcgtcgttgc acctatctag tgctggatga agctgaccgc atgctggaca tgggctttga
     1321 acctcagatc cgcaagatag tggatcaaat acggccagac cgtcagactc tgatgtggag
     1381 tgccacctgg cctaaagaag ttcgtcagct ggctgaggac ttcttgaagg attacgtcca
     1441 gatcaacatt ggcgccctgc agctcagcgc caatcataac atcctgcaaa tagtagatgt
     1501 ttgcaatgac ttagaaaagg aagacaaact gattcggttg ctggaggaga tcatgagtga
     1561 aaaggaaaac aagaccatca tttttgtgga gaccaaaagg cgttgtgatg agctcaccag
     1621 gaggatgaga agagacggat ggcctgcaat gggaatccat ggagataaga gccagcagga
     1681 gagggactgg gtccttaacg agttcagatt tggcaaagct ccgattctca ttgctacaga
     1741 cgtggcctcc cggggcttag atgtggagga tgtgaaattt gtcatcaatt atgactaccc
     1801 taactcctct gaggattata tccaccgcat tggacgcaca gcccgaagtc aaaaaacggg
     1861 cacggcctac accttcttca cccccaacaa catgaaacag gcaggtgacc tgatctctgt
     1921 gctccgtgaa gccaaccagg ccatcaaccc caaactgatc cagatggctg aggacgaccc
     1981 agctttcttg tacaaagtgg ttgactacaa ggatgacgat gacaagctta ccggtgagtt
     2041 cgctatggac aaagattgcg aaatgaaacg taccaccctg gatagcccgc tgggcaaact
     2101 ggaactgagc ggctgcgaac agggcctgca tgaaattaaa ctgctgggta aaggcaccag
     2161 cgcggccgat gcggttgaag ttccggcccc ggccgccgtg ctgggtggtc cggaaccgct
     2221 gatgcaggcg accgcgtggc tgaacgcgta ttttcatcag ccggaagcga ttgaagaatt
     2281 tccggttccg gcgctgcatc atccggtgtt tcagcaggag agctttaccc gtcaggtgct
     2341 gtggaaactg ctgaaagtgg ttaaatttgg cgaagtgatt agctatcagc agctggcggc
     2401 cctggcgggt aatccggcgg ccaccgccgc cgttaaaacc gcgctgagcg gtaacccggt
     2461 gccgattctg attccgtgcc atcgtgtggt tagctctagc ggtgcggttg gcggttatga
     2521 aggtggtctg gcggtgaaag agtggctgct ggcccatgaa ggtcatcgtc tgggtaaacc
     2581 gggtctggga cctgcaggct gataaggatt cgaggctgct aacaaagccc gaaaggaagc
     2641 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     2701 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatccac aggacgggtg
     2761 tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc aggactgggc
     2821 ggcggccaaa gcggtcggac agtgctccga gaacgggtgc gcatagaaat tgcatcaacg
     2881 catatagcgc tagcagcacg ccatagtgac tggcgatgct gtcggaatgg acgatatccc
     2941 gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg
     3001 tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc
     3061 aatttaactg tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag
     3121 aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3181 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3241 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3301 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3361 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3421 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3481 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3541 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     3601 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3661 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3721 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3781 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3841 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     3901 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3961 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4021 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4081 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4141 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4201 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4261 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4321 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4381 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4441 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4501 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4561 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4621 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4681 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4741 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4801 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4861 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4921 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4981 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5041 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5101 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     5161 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
     5221 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     5281 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5341 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5401 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     5461 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     5521 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     5581 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     5641 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     5701 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     5761 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     5821 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     5881 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     5941 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     6001 cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat
     6061 gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt
     6121 ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc
     6181 gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga
     6241 cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc
     6301 catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt
     6361 aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc
     6421 ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata
     6481 atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg
     6541 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
     6601 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     6661 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     6721 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
     6781 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg atcgacgctc
     6841 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     6901 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     6961 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     7021 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     7081 gccggccacg atgcgtccgg cgtagaggat cg
//
