LOCUS       sequence-436837-        8879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DDX3X(G325E) protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8879)
  TITLE     Protein purification and mRNA synthesis plasmids
REFERENCE   2  (bases 1 to 8879)
  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"
COMMENT     Sequence Label: pOCC120-MBP-DDX3X(G325E)-monoGFP-His
FEATURES             Location/Qualifiers
     source          1..8879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        79..170
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     95..112
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      212..221
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             224..1324
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1298..1321
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1379..1402
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3437..3439,3440..3442,3443..4153)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3482..3503)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3743..3762)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4090..4111
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4154..4177
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4187..4204
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4346..4480
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4383..4402)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4437..4456
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           4553..4925
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(5277..5294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5387..5975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5467..5486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6146..6937,6938..7006))
                     /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     6769..6788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7007..7111)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7412..7434
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7426..7443
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7427..7443
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7454..7473
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7454..7472
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             8083..8207
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(8245..8850)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
ORIGIN
        1 agctgccacc attttgttcg cccaggactc tagctatagt tctagtggtt ggctacgtat
       61 actccggaat attaatagat catggagata attaaaatga taaccatctc gcaaataaat
      121 aagtatttta ctgttttcgt aacagttttg taataaaaaa acctataaat attccggatt
      181 attcataccg tcccaccatc gggcgcggat cgccaccatg ggcatgaaaa tcgaagaagg
      241 taaactggta atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa
      301 gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata aactggaaga
      361 gaaattccca caggttgcgg caactggcga tggccctgac attatcttct gggcacacga
      421 ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt
      481 ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc
      541 ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc
      601 gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag
      661 cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg
      721 gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa
      781 cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa
      841 tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac
      901 catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac
      961 ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg
     1021 tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct
     1081 gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa
     1141 gtcttacgag gaagagttgg tgaaagatcc gcgtattgcc gccactatgg aaaacgccca
     1201 gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac
     1261 tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca
     1321 gactaattcg agctcgaaca acaacaacaa taacaataac aacaacagca gcggcagact
     1381 ggaagttctg ttccaggggc cggcggccgc aatgtcccac gtggcagttg agaacgcttt
     1441 gggcctggat caacagttcg ccggattgga cctcaactcg tcggacaacc aatcaggagg
     1501 ctccacggcc tcaaagggac gttacatccc gccacatctc cgcaataggg aggcaaccaa
     1561 aggcttctac gataaagatt cctcaggttg gtcgtcatca aaggataaag atgcttactc
     1621 atcgtttggc tcgaggtcag attcacgcgg caaatccagc ttcttcagtg atcgcggatc
     1681 aggctcacgc ggaaggtttg acgacagggg acgttccgac tatgatggca taggttcccg
     1741 cggtgatcgc agtggctttg gcaaattcga aagaggtggc aattcgcgct ggtgtgacaa
     1801 atctgatgaa gacgactggt cgaaaccact tccgccatct gagagactgg agcaagagct
     1861 gttctcagga ggaaacaccg gaattaattt cgaaaagtac gacgatattc cagtagaagc
     1921 gacgggaaat aactgccctc cacacatcga aagctttagt gacgtcgaaa tgggagagat
     1981 catcatgggt aatatcgaac tcaccagata cactcgtcct actcctgttc aaaaacatgc
     2041 tattccaatt ataaaagaaa agcgtgatct tatggcatgt gcacaaacag gatctggtaa
     2101 gacagcagcc tttctcctgc caatcctttc acagatatac tcggatggcc ccggggaagc
     2161 tctccgtgct atgaaggaaa acggaaggta cggtcgcaga aaacagtatc ctatttcttt
     2221 ggtcctggcc cctacgaggg aactggcggt gcaaatttat gaggaagccc gcaaattttc
     2281 atataggtct agagttaggc cctgtgtcgt atacggagga gctgatatcg gacaacaaat
     2341 ccgtgacctt gaaaggggct gtcaccttct tgtcgccaca ccagaacgct tggtagatat
     2401 gatggaaaga ggaaaaatag gcttggactt ctgcaagtac ctcgtcctcg acgaagccga
     2461 ccgtatgctt gatatgggat ttgagcctca gatacgtcgc atagttgagc aagatacaat
     2521 gccgcctaag ggcgtcaggc acactatgat gttctcagcc acttttccga aggagattca
     2581 gatgttggcc cgtgatttcc tggacgaata catattcctc gccgtcggaa gagtcggtag
     2641 tactagtgaa aatatcaccc aaaaagtcgt ttgggttgag gaaagcgaca aacgtagttt
     2701 ccttcttgat ctgttgaatg ctacgggtaa agacagtctt acactggtgt ttgttgagac
     2761 taaaaaggga gctgactccc ttgaagactt tctgtatcat gaaggatatg cttgtacgtc
     2821 catacatgga gatcgctctc aaagggaccg tgaagaggcg ctccatcaat tccgctccgg
     2881 caagagccca atcttggttg ctaccgcagt cgcggctagg ggtttggata ttagcaatgt
     2941 taagcatgtt atcaacttcg atctccccag cgatatagaa gagtatgttc accgcatcgg
     3001 acgcacaggt cgtgtgggca accttggact ggccacgagt ttcttcaacg aaagaaatat
     3061 taatataacg aaagatctgc tcgacctgct tgtcgaagct aaacaggaag taccatcctg
     3121 gcttgaaaat atggcgtatg aacaccatta taaaggctca tcacgtggaa ggtcaaagag
     3181 tagcaggttt tcgggcggct ttggtgccag agactaccgc cagtcatcgg gtgcaagttc
     3241 gagctccttc tcttccagtc gcgcatcatc tagtcgtagc ggtggcggag gtcatggaag
     3301 cagcagggga ttcggcggag gtggctatgg tggcttctat aattcagacg gctacggagg
     3361 aaactacaac tcgcaaggtg tagactggtg gggaaacggc gcgccgggat ccgctggctc
     3421 cgctgctggt tctggcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3481 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     3541 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3601 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     3661 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3721 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     3781 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     3841 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     3901 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     3961 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     4021 gctgcccgac aaccactacc tgagcaccca gtccaaactg agcaaagacc ccaacgagaa
     4081 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     4141 cgagctgtac aagctggaag ttctgttcca ggggccgggc agcagccatc accatcacca
     4201 tcacagtgga taatagaagc ttgtcgagaa gtactagagg atcataatca gccataccac
     4261 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     4321 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     4381 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4441 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgatcactgc ttgagcctag
     4501 gagatccgaa ccagataagt gaaagcctgc atccgcagat cttaattaat aaaacacgat
     4561 acattgttat tagtacattt attaagcgct agattctgtg cgttgttgat ttacagacaa
     4621 ttgttgtacg tattttaata attcattaaa tttataatct ttagggtggt atgttagagc
     4681 gaaaatcaaa tgattttcag cgtctttata tctgaattta aatattaaat cctcaataga
     4741 tttgtaaaat aggtttcgat tagtttcaaa caagggttgt ttttccgaac cgatggctgg
     4801 actatctaat ggattttcgc tcaacgccac aaaacttgcc aaatcttgta gcagcaatct
     4861 agctttgtcg atattcgttt gtgttttgtt ttgtaataaa ggttcgacgt cgttcaaaat
     4921 attatgcgct tttgtatttc tttcatcact gtcgttagtg tacaattgac tcgacgtaaa
     4981 cacgttaaat aacgcttgga catatttaac atcgggcgtg ttagctttat taggccgatt
     5041 atcgtcgtcg tcccaaccct cgtcgttaga agttgcttcc gaagacgatt ttgccatagc
     5101 cacacgacgc ctattaattg tgtcggctaa cacgtccgcg atcaaatttg tagttgagct
     5161 ttttgggcct catgggcctt ccgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5221 cagctgcatt aacatggtca tagctgtttc cttgcgtatt gggcgctctc cgcttcctcg
     5281 ctcactgact cgctgcgctc ggtcgttcgg gtaaagcctg gggtgcctaa tgagcaaaag
     5341 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5401 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5461 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5521 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5581 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5641 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5701 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5761 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5821 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5881 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5941 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6001 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6061 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6121 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6181 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6241 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagaa ccacgctcac
     6301 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6361 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6421 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6481 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6541 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6601 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6661 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6721 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6781 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6841 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6901 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6961 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7021 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7081 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     7141 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     7201 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     7261 gttgagtggc cgctacaggg cgctcccatt cgccattcag gctgcgcaac tgttgggaag
     7321 ggcgtttcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     7381 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     7441 agtgagcgcg acgtaatacg actcactata gggcgaattg gcggaaggcc gtcaaggccg
     7501 tgacgttaaa actattaagc catccaatcg accgttagtc gaatcaggac cgctggtgcg
     7561 agaagccgcg aagtatggcg aatgcatcgt ataacgtgtg gagtccgctc attagagcgt
     7621 catgtttaga caagaaagct acatatttaa ttgatcccga tgattttatt gataaattga
     7681 ccctaactcc atacacggta ttctacaatg gcggggtttt ggtcaaaatt tccggactgc
     7741 gattgtacat gctgttaacg gctccgccca ctattaatga aattaaaaat tccaatttta
     7801 aaaaacgcag caagagaaac atttgtatga aagaatgcgt agaaggaaag aaaaatgtcg
     7861 tggacatgct gaacaacaag attaatatgc ctccgtgtat aaaaaaaata ttgaacgatt
     7921 tgaaagaaaa caatgtaccg cgcggcggta tgtacaggaa gaggtttata ctaaactgtt
     7981 acattgcaaa cgtggtttcg tgtgccaagt gtgaaaaccg atgtttaatc aaggctctga
     8041 cgcatttcta caaccacgac tccaagtgtg tgggtgaagt catgcatctt ttaatcaaat
     8101 cccaagatgt gtataaacca ccaaactgcc aaaaaatgaa aactgtggac aagctctgtc
     8161 cgtttgctgg caactgcaag ggtctcaatc ctatttgtaa ttattgaata ataaaacaat
     8221 tataaatgct aaatttgttt tttattaacg atacaaacca aacgcaacaa gaacatttgt
     8281 agtattatct ataattgaaa acgcgtagtt ataatcgctg aggtaatatt taaaatcatt
     8341 ttcaaatgat tcacagttaa tttgcgacaa tataatttta ttttcacata aactagacgc
     8401 cttgtcgtct tcttcttcgt attccttctc tttttcattt ttctcctcat aaaaattaac
     8461 atagttatta tcgtatccat atatgtatct atcgtataga gtaaattttt tgttgtcata
     8521 aatatatatg tcttttttaa tggggtgtat agtaccgctg cgcatagttt ttctgtaatt
     8581 tacaacagtg ctattttctg gtagttcttc ggagtgtgtt gctttaatta ttaaatttat
     8641 ataatcaatg aatttgggat cgtcggtttt gtacaatatg ttgccggcat agtacgcagc
     8701 ttcttctagt tcaattacac cattttttag cagcaccgga ttaacataac tttccaaaat
     8761 gttgtacgaa ccgttaaaca aaaacagttc acctcccttt tctatactat tgtctgcgag
     8821 cagttgtttg ttgttaaaaa taacagccat tgtaatgaga cgcacaaact aatatcaca
//
