LOCUS       sequence-266937-        7229 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human DDX43 mutant (K292A, ATPase dead) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7229)
  AUTHORS   Talwar T, Vidhyasagar V, Qing J, Guo M, Kariem A, Lu Y, Singh RS, 
            Lukong KE, Wu Y
  TITLE     The DEAD-box protein DDX43 (HAGE) is a dual RNA-DNA helicase and has
            a K-homology domain required for full nucleic acid unwinding 
            activity.
  JOURNAL   J Biol Chem. 2017 Jun 23;292(25):10429-10443. doi: 
            10.1074/jbc.M117.774950. Epub 2017 May 3.
  PUBMED    28468824
REFERENCE   2  (bases 1 to 7229)
  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: 
            "10.1074/jbc.M117.774950"; journalName: "J Biol Chem"; date: 
            "2017-06-23- 23"; volume: "292"; issue: "25"; pages: "10429-10443"
COMMENT     Sequence Label: pET28a-DDX43-K292A
FEATURES             Location/Qualifiers
     source          1..7229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..466
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     308..329
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(558..1373)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(558..1373)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1281..1300
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1365..1384
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1495..2083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1984..2003
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2237..2254
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2269..2408
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2334..2353)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2494..2516
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2510..2701)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3510..4592)
                     /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     4554..4573
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4593..4670)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4876..4895
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4979..4998
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4979..4997
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4998..5022
                     /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)."
     RBS             5037..5059
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5078..5095
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5105..5122
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(7143..7161)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7157..7204
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaac ttgattaggg tgatggttca
      241 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
      301 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
      361 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
      421 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
      481 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      541 cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc
      601 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
      661 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
      721 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
      781 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
      841 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
      901 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
      961 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     1021 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     1081 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     1141 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     1201 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     1261 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     1321 ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc
     1381 cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa tcccttaacg
     1441 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     1501 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     1561 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1621 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     1681 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     1741 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     1801 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     1861 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     1921 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     1981 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2041 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     2101 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     2161 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     2221 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     2281 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     2341 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     2401 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     2461 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     2521 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     2581 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     2641 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     2701 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     2761 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     2821 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     2881 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
     2941 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     3001 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     3061 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     3121 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc
     3181 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     3241 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     3301 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     3361 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     3421 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     3481 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3541 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3601 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     3661 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     3721 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     3781 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     3841 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     3901 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     3961 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     4021 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     4081 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     4141 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     4201 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     4261 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     4321 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     4381 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     4441 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     4501 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     4561 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     4621 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     4681 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     4741 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     4801 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     4861 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     4921 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaatta
     4981 atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg
     5041 tttaacttta agaaggagat ataccatggg cagcagccat catcatcatc atcacagcag
     5101 cggcctggtg ccgcgcggca gccatatgtc ccaccacgga ggagctccca aggcctctac
     5161 gtgggtcgtt gctagtcggc gaagctcgac agtgtcccga gcgccagaga ggaggccggc
     5221 ggaggagttg aatcgaacag gtcctgaggg atatagtgtc ggcagaggtg gtcgctggag
     5281 aggcacctct aggcccccgg aggacgtggc cgctggtcac gaggaactgc cgctgtgttt
     5341 tgctttgaag agccactttg ttggcgcggt aatcggtcgt ggtgggtcaa aaataaagaa
     5401 tatacaaagt acaacaaaca ccacaatcca aataatacaa gaacaaccag aatcattagt
     5461 caaaattttt ggcagcaagg caatgcaaac gaaagcaaaa gcagtgatag acaattttgt
     5521 taaaaagcta gaagaaaatt acaattcaga atgcggaatt gatactgcat tccaaccttc
     5581 tgttggaaaa gatggaagca cagataacaa tgttgttgca ggagatcggc cattgataga
     5641 ttgggatcaa attagagagg aaggtttgaa atggcaaaaa acaaagtggg cagatttacc
     5701 accaattaag aaaaactttt ataaagagtc cactgccaca agtgccatgt caaaagtaga
     5761 agcagatagt tggaggaaag aaaattttaa tataacgtgg gatgacttga aggatgggga
     5821 gaaacgacct atccccaatc ctacctgcac atttgatgac gcctttcaat gttatcctga
     5881 ggttatggaa aacattaaaa aggcaggttt tcaaaagcca acacctattc agtcacaggc
     5941 atggcccatt gtgttgcaag gaatagatct tataggagta gcccagactg gaacaggagc
     6001 gacattgtgt tatttaatgc ctggatttat tcatctggtc cttcaaccca gccttaaagg
     6061 tcaaaggaat agacccggca tgttagttct aactcccact cgggaattag cacttcaagt
     6121 agaaggagaa tgttgcaaat attcatataa agggcttcgg agtgtttgtg tatatggtgg
     6181 tggaaataga gatgaacaaa tagaagagct taaaaaaggt gtagatatca taattgcaac
     6241 tcccggaaga ttgaatgatc tgcaaatgag taacttcgtc aatctgaaga atataaccta
     6301 cttggtttta gatgaagcag acaagatgtt ggacatggga tttgaacccc agataatgaa
     6361 gattttgtta gatgtgcgcc cagataggca gacagttatg accagtgcta catggcctca
     6421 ttcagttcat cgcctcgcac aatcttattt gaaagaacca atgattgtct atgttggtac
     6481 attggatcta gttgctgtaa gttcagtgaa gcaaaatata attgtaacca ccgaggaaga
     6541 gaaatggagt cacatgcaaa cttttctaca gagtatgtca tccacagaca aagtcattgt
     6601 cttcgtttct cgaaaagctg ttgcggatca cttatcaagt gacctaatac ttggaaatat
     6661 atcagtagag tctctgcatg gagatagaga acagagagat cgggagaaag cattagagaa
     6721 ctttaaaaca ggcaaagtga gaatactaat tgcaactgat ctagcctcta gaggacttga
     6781 tgtccatgac gttacacatg tctataattt tgactttcca cggaatattg aagaatacgt
     6841 acaccgaata gggcgcacgg gaagagcagg gaggactggt gtttccatta caactttgac
     6901 tagaaatgat tggagggttg cctctgaatt gattaatatt ctggaaagag caaatcagag
     6961 tattccagag gagcttgtat caatggctga gaggtttaag gcacatcaac agaaaaggga
     7021 aatggaaaga aaaatggaaa gacctcaagg aaggcccaag aagtttcatc tcgagcacca
     7081 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     7141 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     7201 tttgctgaaa ggaggaacta tatccggat
//
