LOCUS       sequence-441381-        6589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express DDX53 protein in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6589)
  TITLE     DDX53 and 3 x FLAG
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6589)
  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            "Unpublished"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: pDEST17-DDX53
FEATURES             Location/Qualifiers
     source          1..6589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             113..130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    140..164
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2058..2082)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2135..2153)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2149..2196
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2562..2567,2568..2584,2585..2590))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2617..2635)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2703..2807
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2808..2876,2877..3668)
                     /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(3026..3045)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3839..4427
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4328..4347
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4581..4598
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4613..4752
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4678..4697)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4838..4860
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4854..5045)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6507..6526
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagaaataat tttgtttaac tttaagaagg agatatacat atgtcgtact accatcacca
      121 tcaccatcac ctcgaatcaa caagtttgta caaaaaagtt ggcatgtccc actgggcccc
      181 agagtggaag agggcggagg ctaatccaag agaccttggg gccagctggg atgtcagggg
      241 cagcagaggc agtggctgga gtggcccctt cggccatcag ggaccgagag cagcaggctc
      301 ccgtgaacca ccactctgct ttaaaataaa gaacaatatg gttggtgtgg tcattggtta
      361 cagtggatca aaaataaaag atctacaaca ttcgacaaac actaaaatac agatcataaa
      421 cggggaatct gaagcaaaag tcagaatttt tggcaatagg gaaatgaaag caaaggccaa
      481 agcggctata gaaacactta ttagaaaaca agaaagctac aactcagaat ccagtgtgga
      541 taatgctgca tcccaaaccc ctattggaag aaatctaggc agaaatgaca ttgttggaga
      601 agctgagcca ttgtcaaatt gggatcgcat tagggcagca gtcgtggagt gcgaaaagag
      661 aaaatgggca gatctaccac cagttaagaa aaacttttac atagaatcca aagcaacaag
      721 ctgcatgtct gaaatgcagg tgattaactg gagaaaggaa aatttcaaca taacgtgtga
      781 tgacttgaaa agtggtgaaa agcgtctcat tccaaaacca acttgcaggt ttaaagacgc
      841 ttttcagcaa taccctgatc ttctgaaaag cataataagg gtagggattt taaagccaac
      901 gccaattcag tcacaggcat ggccaattat tctacaagga atagatctta tagtagttgc
      961 acaaaccgga acagggaaaa cattgtccta tctaatgcct gggtttattc atcttgattc
     1021 tcaaccaata tctagagagc aaaggaatgg gcctgggatg ctagtcctta cacccactag
     1081 agagttggct cttcacgtgg aagctgaatg ttcaaagtat tcatataaag gtctcaaaag
     1141 catttgcata tatggtggta gaaacagaaa tggacaaata gaagacatta gcaaaggtgt
     1201 agatatcatt attgcaactc ctgggaggct gaatgaccta caaatgaata actctgtcaa
     1261 cctaagaagc ataacctact tggttataga tgaggcagat aaaatgctgg atatggaatt
     1321 tgaaccccag ataaggaaga ttttattaga tgtgcgccca gaccgacaga ctgttatgac
     1381 aagtgcaact tggccagata ctgtacgtca actagcactt tcttatttga aagatcctat
     1441 gattgtttat gttggtaatc tgaatctagt ggctgtaaat acagtgaagc aaaatataat
     1501 tgttaccaca gaaaaagaaa aacgagctct cacccaagaa ttcgtagaga acatgtcacc
     1561 caacgacaaa gtcatcatgt ttgtcagcca aaaacatatt gctgatgact tgtcaagcga
     1621 cttcaatatc caaggcatat ctgcagaatc attacatggc aacagtgaac agagtgatca
     1681 agagcgagca gtagaggact ttaaaagcgg aaacataaag atactgatta caactgatat
     1741 agtatcccga ggtcttgatc ttaatgatgt cacacatgta tataattatg atttcccaag
     1801 gaatattgac gtatatgtac acagagtagg gtacattgga cggacaggaa agactggcac
     1861 atcagttacc ctcatcactc agagagattc gaaaatggcc ggtgaattga ttaaaattct
     1921 ggacagagca aatcagagtg ttccggaaga tcttgtagta atggctgagc aatacaagtt
     1981 aaatcaacaa aagaggcaca gagaaacacg atcaagagaa cctggacaaa gacgtaagga
     2041 gttttatttt ttaagttgcc caactttctt gtacaaagtg gttgattcga ggctgctaac
     2101 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     2161 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     2221 tatccacagg acgggtgtgg tcgccatgat cgcgtagtcg atagtggctc caagtagcga
     2281 agcgagcagg actgggcggc ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca
     2341 tagaaattgc atcaacgcat atagcgctag cagcacgcca tagtgactgg cgatgctgtc
     2401 ggaatggacg atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac
     2461 agcatccagg gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg
     2521 tgcctgactg cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat
     2581 aagctgtcaa acatgagaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat
     2641 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     2701 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2761 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2821 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2881 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2941 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3001 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     3061 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3121 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3181 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3241 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3301 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     3361 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3421 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3481 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3541 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3601 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3661 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3721 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3781 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3841 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3901 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3961 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     4021 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4081 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4141 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4201 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4261 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4321 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4381 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4441 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4501 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4561 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     4621 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     4681 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     4741 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4801 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4861 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     4921 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     4981 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     5041 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     5101 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     5161 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     5221 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     5281 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     5341 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     5401 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     5461 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac
     5521 ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg
     5581 ttctgccaag ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt
     5641 cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc
     5701 ggctccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa
     5761 tccatgccaa cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg
     5821 gtccaatgat cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat
     5881 ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg
     5941 aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga
     6001 cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt
     6061 tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa
     6121 gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga
     6181 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga
     6241 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     6301 tcggtcgatc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     6361 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     6421 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     6481 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     6541 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcg
//
