LOCUS       sequence-181029-        6798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inducible expression of GST-DDX5 D248N in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6798)
  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 6798)
  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: pGEX_hDDX5_FL_D248N
FEATURES             Location/Qualifiers
     source          1..6798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2848..2870)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3015..3033)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3101..3205
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3206..3274,3275..4066)
                     /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(3424..3443)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4237..4825
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4726..4745
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4979..4996
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5069..5146
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5147..6229
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5166..5185)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6278..6283,6284..6301,6302..6308)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6316..6332
                     /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)."
     primer_bind     6340..6356
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6340..6356
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6352..6798,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6372..6389)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6372..6388)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6381..6403)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gttccgcgtg gatcctcggg ttattcgagt gaccgagacc
      961 gcggccggga ccgagggttt ggtgcacctc gatttggagg aagtagggca gggcccttat
     1021 ctggaaagaa gtttggaaac cctggggaga aattagttaa aaagaagtgg aatcttgatg
     1081 agctgcctaa atttgagaag aatttttatc aagagcaccc tgatttggct aggcgcacag
     1141 cacaagaggt ggaaacatac agaagaagca aggaaattac agttagaggt cacaactgcc
     1201 cgaagccagt tctaaatttt tatgaagcca atttccctgc aaatgtcatg gatgttattg
     1261 caagacagaa tttcactgaa cccactgcta ttcaagctca gggatggcca gttgctctaa
     1321 gtggattgga tatggttgga gtggcacaga ctggatctgg gaaaacattg tcttatttgc
     1381 ttcctgccat tgtccacatc aatcatcagc cattcctaga gagaggcgat gggcctattt
     1441 gtttggtgct ggcaccaact cgggaactgg cccaacaggt gcagcaagta gctgctgaat
     1501 attgtagagc atgtcgcttg aagtctactt gtatctacgg tggtgctcct aagggaccac
     1561 aaatacgtga tttggagaga ggtgtggaaa tctgtattgc aacacctgga agactgattg
     1621 actttttaga gtgtggaaaa accaatctga gaagaacaac ctaccttgtc cttaatgaag
     1681 cagatagaat gcttgatatg ggctttgaac cccaaataag gaagattgtg gatcaaataa
     1741 gacctgatag gcaaactcta atgtggagtg cgacttggcc aaaagaagta agacagcttg
     1801 ctgaagattt cctgaaagac tatattcata taaacattgg tgcacttgaa ctgagtgcaa
     1861 accacaacat tcttcagatt gtggatgtgt gtcatgacgt agaaaaggat gaaaaactta
     1921 ttcgtctaat ggaagagatc atgagtgaga aggagaataa aaccattgtt tttgtggaaa
     1981 ccaaaagaag atgtgatgag cttaccagaa aaatgaggag agatgggtgg cctgccatgg
     2041 gtatccatgg tgacaagagt caacaagagc gtgactgggt tctaaatgaa ttcaaacatg
     2101 gaaaagctcc tattctgatt gctacagatg tggcctccag agggctagat gtggaagatg
     2161 tgaaatttgt catcaattat gactacccta actcctcaga ggattatatt catcgaattg
     2221 gaagaactgc tcgcagtacc aaaacaggca cagcatacac tttctttaca cctaataaca
     2281 taaagcaagt gagcgacctt atctctgtgc ttcgtgaagc taatcaagca attaatccca
     2341 agttgcttca gttggtcgaa gacagaggtt caggtcgttc caggggtaga ggaggcatga
     2401 aggatgaccg tcgggacaga tactctgcgg gcaaaagggg tggatttaat acctttagag
     2461 acagggaaaa ttatgacaga ggttactcta gcctgcttaa aagagatttt ggggcaaaaa
     2521 ctcagaatgg tgtttacagt gctgcaaatt acaccaatgg gagctttgga agtaattttg
     2581 tgtctgctgg tatacagacc agttttagga ctggtaatcc aacagggact taccagaatg
     2641 gttatgatag cactcagcaa tacggaagta atgttccaaa tatgcacaat ggtatgaacc
     2701 aacaggcata tgcatatcct gctactgcag ctgcacctat gattggttat ccaatgccaa
     2761 caggatattc ccaataagtc gactcgagcg gccgcatcgt gactgactga cgatctgcct
     2821 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     2881 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     2941 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtataattc
     3001 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3061 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3121 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3181 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3241 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3301 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3361 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3421 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     3481 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3541 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3601 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3661 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3721 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     3781 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3841 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3901 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     3961 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4021 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4081 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4141 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4201 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4261 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4321 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4381 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4441 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4501 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4561 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4621 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4681 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4741 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4801 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4861 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4921 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     4981 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     5041 gtgcggtatt tcacaccgca taaattccga caccatcgaa tggcgcaaaa cctttcgcgg
     5101 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
     5161 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
     5221 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
     5281 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
     5341 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
     5401 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
     5461 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
     5521 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
     5581 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     5641 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
     5701 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
     5761 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
     5821 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
     5881 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
     5941 tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
     6001 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
     6061 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
     6121 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6181 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     6241 ttaatgtaag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     6301 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     6361 attacggatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     6421 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     6481 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg ctttgcctgg
     6541 tttccggcac cagaagcggt gccggaaagc tggctggagt gcgatcttcc tgaggccgat
     6601 actgtcgtcg tcccctcaaa ctggcagatg cacggttacg atgcgcccat ctacaccaac
     6661 gtgacctatc ccattacggt caatccgccg tttgttccca cggagaatcc gacgggttgt
     6721 tactcgctca catttaatgt tgatgaaagc tggctacagg aaggccagac gcgaattatt
     6781 tttgatggcg ttggaatt
//
