LOCUS       sequence-436790-        8111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DDX3X(WT) protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8111)
  TITLE     Protein purification and mRNA synthesis plasmids
REFERENCE   2  (bases 1 to 8111)
  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: pOCC102-His-MBP-DDX3X(WT)
FEATURES             Location/Qualifiers
     source          1..8111
                     /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             230..247
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             260..1360
                     /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     1334..1357
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1415..1438
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    3578..3712
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3615..3634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3669..3688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           3785..4157
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(4509..4526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4619..5207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4699..4718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5378..6169,6170..6238))
                     /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     6001..6020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6239..6343)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6644..6666
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6658..6675
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6659..6675
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6686..6705
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6686..6704
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7315..7439
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(7477..8082)
                     /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 ggcagcagcc atcaccatca
      241 ccatcacagc agcggcagaa tgaaaatcga agaaggtaaa ctggtaatct ggattaacgg
      301 cgataaaggc tataacggtc tcgctgaagt cggtaagaaa ttcgagaaag ataccggaat
      361 taaagtcacc gttgagcatc cggataaact ggaagagaaa ttcccacagg ttgcggcaac
      421 tggcgatggc cctgacatta tcttctgggc acacgaccgc tttggtggct acgctcaatc
      481 tggcctgttg gctgaaatca ccccggacaa agcgttccag gacaagctgt atccgtttac
      541 ctgggatgcc gtacgttaca acggcaagct gattgcttac ccgatcgctg ttgaagcgtt
      601 atcgctgatt tataacaaag atctgctgcc gaacccgcca aaaacctggg aagagatccc
      661 ggcgctggat aaagaactga aagcgaaagg taagagcgcg ctgatgttca acctgcaaga
      721 accgtacttc acctggccgc tgattgctgc tgacgggggt tatgcgttca agtatgaaaa
      781 cggcaagtac gacattaaag acgtgggcgt ggataacgct ggcgcgaaag cgggtctgac
      841 cttcctggtt gacctgatta aaaacaaaca catgaatgca gacaccgatt actccatcgc
      901 agaagctgcc tttaataaag gcgaaacagc gatgaccatc aacggcccgt gggcatggtc
      961 caacatcgac accagcaaag tgaattatgg tgtaacggta ctgccgacct tcaagggtca
     1021 accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt aacgccgcca gtccgaacaa
     1081 agagctggca aaagagttcc tcgaaaacta tctgctgact gatgaaggtc tggaagcggt
     1141 taataaagac aaaccgctgg gtgccgtagc gctgaagtct tacgaggaag agttggtgaa
     1201 agatccgcgt attgccgcca ctatggaaaa cgcccagaaa ggtgaaatca tgccgaacat
     1261 cccgcagatg tccgctttct ggtatgccgt gcgtactgcg gtgatcaacg ccgccagcgg
     1321 tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact aattcgagct cgaacaacaa
     1381 caacaataac aataacaaca acagcagcgg cagactggaa gttctgttcc aggggccggc
     1441 ggccgcaatg tcccacgtgg cagttgagaa cgctttgggc ctggatcaac agttcgccgg
     1501 attggacctc aactcgtcgg acaaccaatc aggaggctcc acggcctcaa agggacgtta
     1561 catcccgcca catctccgca atagggaggc aaccaaaggc ttctacgata aagattcctc
     1621 aggttggtcg tcatcaaagg ataaagatgc ttactcatcg tttggctcga ggtcagattc
     1681 acgcggcaaa tccagcttct tcagtgatcg cggatcaggc tcacgcggaa ggtttgacga
     1741 caggggacgt tccgactatg atggcatagg ttcccgcggt gatcgcagtg gctttggcaa
     1801 attcgaaaga ggtggcaatt cgcgctggtg tgacaaatct gatgaagacg actggtcgaa
     1861 accacttccg ccatctgaga gactggagca agagctgttc tcaggaggaa acaccggaat
     1921 taatttcgaa aagtacgacg atattccagt agaagcgacg ggaaataact gccctccaca
     1981 catcgaaagc tttagtgacg tcgaaatggg agagatcatc atgggtaata tcgaactcac
     2041 cagatacact cgtcctactc ctgttcaaaa acatgctatt ccaattataa aagaaaagcg
     2101 tgatcttatg gcatgtgcac aaacaggatc tggtaagaca gcagcctttc tcctgccaat
     2161 cctttcacag atatactcgg atggccccgg ggaagctctc cgtgctatga aggaaaacgg
     2221 aaggtacggt cgcagaaaac agtatcctat ttctttggtc ctggccccta cgagggaact
     2281 ggcggtgcaa atttatgagg aagcccgcaa attttcatat aggtctagag ttaggccctg
     2341 tgtcgtatac ggaggagctg atatcggaca acaaatccgt gaccttgaaa ggggctgtca
     2401 ccttcttgtc gccacaccag gacgcttggt agatatgatg gaaagaggaa aaataggctt
     2461 ggacttctgc aagtacctcg tcctcgacga agccgaccgt atgcttgata tgggatttga
     2521 gcctcagata cgtcgcatag ttgagcaaga tacaatgccg cctaagggcg tcaggcacac
     2581 tatgatgttc tcagccactt ttccgaagga gattcagatg ttggcccgtg atttcctgga
     2641 cgaatacata ttcctcgccg tcggaagagt cggtagtact agtgaaaata tcacccaaaa
     2701 agtcgtttgg gttgaggaaa gcgacaaacg tagtttcctt cttgatctgt tgaatgctac
     2761 gggtaaagac agtcttacac tggtgtttgt tgagactaaa aagggagctg actcccttga
     2821 agactttctg tatcatgaag gatatgcttg tacgtccata catggagatc gctctcaaag
     2881 ggaccgtgaa gaggcgctcc atcaattccg ctccggcaag agcccaatct tggttgctac
     2941 cgcagtcgcg gctaggggtt tggatattag caatgttaag catgttatca acttcgatct
     3001 ccccagcgat atagaagagt atgttcaccg catcggacgc acaggtcgtg tgggcaacct
     3061 tggactggcc acgagtttct tcaacgaaag aaatattaat ataacgaaag atctgctcga
     3121 cctgcttgtc gaagctaaac aggaagtacc atcctggctt gaaaatatgg cgtatgaaca
     3181 ccattataaa ggctcatcac gtggaaggtc aaagagtagc aggttttcgg gcggctttgg
     3241 tgccagagac taccgccagt catcgggtgc aagttcgagc tccttctctt ccagtcgcgc
     3301 atcatctagt cgtagcggtg gcggaggtca tggaagcagc aggggattcg gcggaggtgg
     3361 ctatggtggc ttctataatt cagacggcta cggaggaaac tacaactcgc aaggtgtaga
     3421 ctggtgggga aacggcgcgc cgtaatagaa gcttgtcgag aagtactaga ggatcataat
     3481 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3541 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3601 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3661 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctgatcact
     3721 gcttgagcct aggagatccg aaccagataa gtgaaagcct gcatccgcag atcttaatta
     3781 ataaaacacg atacattgtt attagtacat ttattaagcg ctagattctg tgcgttgttg
     3841 atttacagac aattgttgta cgtattttaa taattcatta aatttataat ctttagggtg
     3901 gtatgttaga gcgaaaatca aatgattttc agcgtcttta tatctgaatt taaatattaa
     3961 atcctcaata gatttgtaaa ataggtttcg attagtttca aacaagggtt gtttttccga
     4021 accgatggct ggactatcta atggattttc gctcaacgcc acaaaacttg ccaaatcttg
     4081 tagcagcaat ctagctttgt cgatattcgt ttgtgttttg ttttgtaata aaggttcgac
     4141 gtcgttcaaa atattatgcg cttttgtatt tctttcatca ctgtcgttag tgtacaattg
     4201 actcgacgta aacacgttaa ataacgcttg gacatattta acatcgggcg tgttagcttt
     4261 attaggccga ttatcgtcgt cgtcccaacc ctcgtcgtta gaagttgctt ccgaagacga
     4321 ttttgccata gccacacgac gcctattaat tgtgtcggct aacacgtccg cgatcaaatt
     4381 tgtagttgag ctttttgggc ctcatgggcc ttccgctcac tgcccgcttt ccagtcggga
     4441 aacctgtcgt gccagctgca ttaacatggt catagctgtt tccttgcgta ttgggcgctc
     4501 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc gggtaaagcc tggggtgcct
     4561 aatgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4621 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4681 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4741 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4801 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4861 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4921 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4981 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5041 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5101 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5161 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5221 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5281 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5341 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5401 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5461 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5521 aaccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5581 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5641 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5701 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5761 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5821 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5881 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5941 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6001 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6061 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6121 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6181 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6241 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6301 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6361 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     6421 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     6481 gaccgagata gggttgagtg gccgctacag ggcgctccca ttcgccattc aggctgcgca
     6541 actgttggga agggcgtttc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     6601 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     6661 aaaacgacgg ccagtgagcg cgacgtaata cgactcacta tagggcgaat tggcggaagg
     6721 ccgtcaaggc cgtgacgtta aaactattaa gccatccaat cgaccgttag tcgaatcagg
     6781 accgctggtg cgagaagccg cgaagtatgg cgaatgcatc gtataacgtg tggagtccgc
     6841 tcattagagc gtcatgttta gacaagaaag ctacatattt aattgatccc gatgatttta
     6901 ttgataaatt gaccctaact ccatacacgg tattctacaa tggcggggtt ttggtcaaaa
     6961 tttccggact gcgattgtac atgctgttaa cggctccgcc cactattaat gaaattaaaa
     7021 attccaattt taaaaaacgc agcaagagaa acatttgtat gaaagaatgc gtagaaggaa
     7081 agaaaaatgt cgtggacatg ctgaacaaca agattaatat gcctccgtgt ataaaaaaaa
     7141 tattgaacga tttgaaagaa aacaatgtac cgcgcggcgg tatgtacagg aagaggttta
     7201 tactaaactg ttacattgca aacgtggttt cgtgtgccaa gtgtgaaaac cgatgtttaa
     7261 tcaaggctct gacgcatttc tacaaccacg actccaagtg tgtgggtgaa gtcatgcatc
     7321 ttttaatcaa atcccaagat gtgtataaac caccaaactg ccaaaaaatg aaaactgtgg
     7381 acaagctctg tccgtttgct ggcaactgca agggtctcaa tcctatttgt aattattgaa
     7441 taataaaaca attataaatg ctaaatttgt tttttattaa cgatacaaac caaacgcaac
     7501 aagaacattt gtagtattat ctataattga aaacgcgtag ttataatcgc tgaggtaata
     7561 tttaaaatca ttttcaaatg attcacagtt aatttgcgac aatataattt tattttcaca
     7621 taaactagac gccttgtcgt cttcttcttc gtattccttc tctttttcat ttttctcctc
     7681 ataaaaatta acatagttat tatcgtatcc atatatgtat ctatcgtata gagtaaattt
     7741 tttgttgtca taaatatata tgtctttttt aatggggtgt atagtaccgc tgcgcatagt
     7801 ttttctgtaa tttacaacag tgctattttc tggtagttct tcggagtgtg ttgctttaat
     7861 tattaaattt atataatcaa tgaatttggg atcgtcggtt ttgtacaata tgttgccggc
     7921 atagtacgca gcttcttcta gttcaattac accatttttt agcagcaccg gattaacata
     7981 actttccaaa atgttgtacg aaccgttaaa caaaaacagt tcacctccct tttctatact
     8041 attgtctgcg agcagttgtt tgttgttaaa aataacagcc attgtaatga gacgcacaaa
     8101 ctaatatcac a
//
