LOCUS       sequence-181024-        7330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of Ha-DDX5 K144N in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7330)
  AUTHORS   Huang W, Thomas B, Flynn RA, Gavzy SJ, Wu L, Kim SV, Hall JA, 
            Miraldi ER, Ng CP, Rigo F, Meadows S, Montoya NR, Herrera NG, 
            Domingos AI, Rastinejad F, Myers RM, Fuller-Pace FV, Bonneau R, 
            Chang HY, Acuto O, Littman DR
  TITLE     DDX5 and its associated lncRNA Rmrp modulate TH17 cell effector 
            functions.
  JOURNAL   Nature. 2015 Dec 24;528(7583):517-22. doi: 10.1038/nature16193. Epub
            2015 Dec 16.
  PUBMED    26675721
REFERENCE   2  (bases 1 to 7330)
  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: 
            "10.1038/nature16193"; journalName: "Nature"; date: "2015-12-24- 
            24"; volume: "528"; issue: "7583"; pages: "517-22"
COMMENT     Sequence Label: pcDHa_mDDX5_FL_K144N
FEATURES             Location/Qualifiers
     source          1..7330
                     /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"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             677..703
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2649..2667)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2650..2667)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2687..2704)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2693..2917
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2963..3391
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3050..3069)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3260..3281
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3400..3420)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3405..3734
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3585..3720
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3647..3666
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3801..4595
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3855..3874)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4465..4484
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4769..4890
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4806..4825)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4860..4879
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4939..4955)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4939..4955)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4952..4974)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4963..4979
                     /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(4987..4993,4994..5011,5012..5017))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5032..5053
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5170..5187)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5341..5929)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5421..5440)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6100..6891,6892..6960))
                     /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     6723..6742
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6961..7065)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7140..7159)
                     /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 agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 gaccaccatg ggtacctacc cttatgacgt ccccgattac gccggatcca ctagtaacgg
      721 ccgccagtgt gctggaattc tgcagatatc catcacactg gcggccgctt cgagttattc
      781 tagtgaccga gaccgcggcc gggatcgagg gtttggtgca cctcgatttg gagggagtag
      841 aacaggaccc ctctctggaa agaagtttgg aaatcctggg gagaaactag ttaaaaagaa
      901 gtggaatctt gatgagctgc ccaaatttga gaagaatttt tatcaagaac accctgattt
      961 ggcaaggcgc accgcacaag aggtagatac atacagaaga agcaaggaaa ttacagttag
     1021 aggtcacaac tgtccaaaac ctgttctgaa tttttatgaa gcaaactttc ctgcgaatgt
     1081 catggatgtg attgcaaggc agaactttac tgaacccact gctattcaag ctcagggctg
     1141 gccagttgct ctcagtggat tggatatggt tggagtagct cagactggat ctgggaacac
     1201 attatcttat ttgctgcctg ccattgtaca cataaaccac cagccattcc tagagagagg
     1261 tgatgggcct atttgcttgg tgctggcacc aactcgagaa ctggcacagc aggtgcagca
     1321 agtggctgct gaatattgtc gagcttgtcg cttgaagtct acttgcatct atggtggtgc
     1381 tcccaaagga ccacagattc gtgatttgga aagaggtgtg gaaatctgta ttgcaacacc
     1441 tggaagactg attgactttt tagagtgtgg gaaaaccaat ctgagaagaa caacttacct
     1501 tgtccttgat gaagctgata ggatgcttga tatgggattt gaaccccaga taaggaaaat
     1561 tgtggatcaa ataagacctg ataggcaaac actaatgtgg agtgcaactt ggccaaaaga
     1621 agtaagacag cttgctgaag atttcctgaa agactatatt catatcaata ttggtgcact
     1681 ggaactgagt gcaaaccata acattcttca gattgtggat gtatgtcatg atgtcgaaaa
     1741 ggatgaaaag cttattcgtc tgatggaaga aatcatgagt gagaaggaga ataaaactat
     1801 tgtttttgtt gaaaccaaaa gaagatgtga tgaacttacc agaaaaatga ggagagatgg
     1861 gtggcctgcc atgggcatcc atggtgacaa gagtcagcag gaacgtgact gggttctaaa
     1921 tgaattcaaa catggaaaag ctcctattct gattgctacc gatgtggcct ccagagggct
     1981 agatgtggaa gatgtgaaat ttgtcatcaa ttatgactac cctaactcct cagaggatta
     2041 tattcatcga attggaagaa ctgctcgcag taccaaaaca ggcacagcat acactttctt
     2101 tacacctaat aacataaagc aagtgagcga ccttatctct gtgcttcggg aagctaatca
     2161 agcaattaat cccaagttgc ttcagttggt cgaagacaga ggttcaggtc gttccagggg
     2221 tagaggaggc atgaaggacg atcgtcgtga cagatactct gcaggcaaaa ggggtggatt
     2281 taataccttt agagacaggg aaaactatga cagaggctac tctaatctgc ttaagagaga
     2341 ttttggggct aaaactcaga atggtgttta cagtgctgca aattacacca atgggagctt
     2401 tggaagtaat tttgtatctg ctggcataca gaccagtttt aggactggta atccaacagg
     2461 gacttaccag aacggttatg atagcactca gcaatatgga agtaatgttg caaatatgca
     2521 caatggtatg aaccaacagg catatgcata tcctgctacc gcagctgctg cgcctatgat
     2581 tggctatccc atgccaacag ggtattctca ataagcggcc gctcgagcat gcatctagag
     2641 ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct
     2701 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2761 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2821 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2881 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2941 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3001 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3061 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3121 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3181 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3241 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3301 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3361 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3421 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3481 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3541 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3601 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3661 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3721 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3781 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3841 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3901 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3961 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4021 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4081 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4141 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4201 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4261 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4321 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4381 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4441 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4501 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4561 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4621 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4681 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4741 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4801 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4861 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4921 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4981 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5041 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5101 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5161 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5221 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5281 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5341 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5401 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5461 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5521 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5581 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5641 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5701 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5761 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5821 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5881 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5941 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6001 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6061 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6121 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6181 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6241 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6301 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6361 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6421 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6481 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6541 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6601 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6661 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6721 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6781 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6841 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6901 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6961 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7021 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7081 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     7141 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     7201 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     7261 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     7321 tatacgcgtt
//
