LOCUS       sequence-441373-        7340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FLAG tagged DDX53 mutant K272A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7340)
  TITLE     DDX53 and 3 x FLAG
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7340)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA-DDX53-K272A
FEATURES             Location/Qualifiers
     source          1..7340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2797..2862
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(2893..2911)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2894..2911)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2931..2948)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2937..3161
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3207..3635
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3294..3313)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3504..3525
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3644..3664)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3649..3978
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3829..3964
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3891..3910
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4045..4839
                     /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(4099..4118)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4709..4728
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5013..5134
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5050..5069)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5104..5123
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5183..5199)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5183..5199)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5196..5218)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5207..5223
                     /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(5231..5237,5238..5255,5256..5261))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5276..5297
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5414..5431)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5585..6173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5665..5684)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6344..7135,7136..7204))
                     /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     6967..6986
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7205..7309)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggcggc
      901 accatgtccc actgggcccc agagtggaag agggcggagg ctaatccaag agaccttggg
      961 gccagctggg atgtcagggg cagcagaggc agtggctgga gtggcccctt cggccatcag
     1021 ggaccgagag cagcaggctc ccgtgaacca ccactctgct ttaaaataaa gaacaatatg
     1081 gttggtgtgg tcattggtta cagtggatca aaaataaaag atctacaaca ttcgacaaac
     1141 actaaaatac agatcataaa cggggaatct gaagcaaaag tcagaatttt tggcaatagg
     1201 gaaatgaaag caaaggccaa agcggctata gaaacactta ttagaaaaca agaaagctac
     1261 aactcagaat ccagtgtgga taatgctgca tcccaaaccc ctattggaag aaatctaggc
     1321 agaaatgaca ttgttggaga agctgagcca ttgtcaaatt gggatcgcat tagggcagca
     1381 gtcgtggagt gcgaaaagag aaaatgggca gatctaccac cagttaagaa aaacttttac
     1441 atagaatcca aagcaacaag ctgcatgtct gaaatgcagg tgattaactg gagaaaggaa
     1501 aatttcaaca taacgtgtga tgacttgaaa agtggtgaaa agcgtctcat tccaaaacca
     1561 acttgcaggt ttaaagacgc ttttcagcaa taccctgatc ttctgaaaag cataataagg
     1621 gtagggattt taaagccaac gccaattcag tcacaggcat ggccaattat tctacaagga
     1681 atagatctta tagtagttgc acaaaccgga acaggggcaa cattgtccta tctaatgcct
     1741 gggtttattc atcttgattc tcaaccaata tctagagagc aaaggaatgg gcctgggatg
     1801 ctagtcctta cacccactag agagttggct cttcacgtgg aagctgaatg ttcaaagtat
     1861 tcatataaag gtctcaaaag catttgcata tatggtggta gaaacagaaa tggacaaata
     1921 gaagacatta gcaaaggtgt agatatcatt attgcaactc ctgggaggct gaatgaccta
     1981 caaatgaata actctgtcaa cctaagaagc ataacctact tggttataga tgaggcagat
     2041 aaaatgctgg atatggaatt tgaaccccag ataaggaaga ttttattaga tgtgcgccca
     2101 gaccgacaga ctgttatgac aagtgcaact tggccagata ctgtacgtca actagcactt
     2161 tcttatttga aagatcctat gattgtttat gttggtaatc tgaatctagt ggctgtaaat
     2221 acagtgaagc aaaatataat tgttaccaca gaaaaagaaa aacgagctct cacccaagaa
     2281 ttcgtagaga acatgtcacc caacgacaaa gtcatcatgt ttgtcagcca aaaacatatt
     2341 gctgatgact tgtcaagcga cttcaatatc caaggcatat ctgcagaatc attacatggc
     2401 aacagtgaac agagtgatca agagcgagca gtagaggact ttaaaagcgg aaacataaag
     2461 atactgatta caactgatat agtatcccga ggtcttgatc ttaatgatgt cacacatgta
     2521 tataattatg atttcccaag gaatattgac gtatatgtac acagagtagg gtacattgga
     2581 cggacaggaa agactggcac atcagttacc ctcatcactc agagagattc gaaaatggcc
     2641 ggtgaattga ttaaaattct ggacagagca aatcagagtg ttccggaaga tcttgtagta
     2701 atggctgagc aatacaagtt aaatcaacaa aagaggcaca gagaaacacg atcaagagaa
     2761 cctggacaaa gacgtaagga gttttatttt ttaagtgact acaaagacca tgacggtgat
     2821 tataaagatc atgacatcga ttacaaggat gacgatgaca agtgactcga gcatgcatct
     2881 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
     2941 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3001 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3061 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3121 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3181 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3241 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3301 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3361 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3421 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3481 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3541 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3601 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3661 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3721 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3781 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3841 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3901 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3961 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4021 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4081 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4141 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4201 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4261 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4321 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4381 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4441 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4501 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4561 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4621 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4681 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4741 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4801 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4861 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4921 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4981 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5041 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5101 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5161 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5221 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5281 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5341 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5401 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5461 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5521 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5581 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5641 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5701 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5761 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5821 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5881 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5941 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6001 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6061 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6121 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6181 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6241 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6301 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6361 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6421 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6481 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6541 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6601 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6661 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6721 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6781 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6841 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6901 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6961 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7021 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7081 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7141 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7201 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7261 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7321 gaaaagtgcc acctgacgtc
//
