LOCUS       sequence-450089-        5890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For purification of N-terminally His-tagged DXO from bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5890)
  AUTHORS   Weixler L, Ikenga NJ, Voorneveld J, Aydin G, Bolte TM, Momoh J, 
            Butepage M, Golzmann A, Luscher B, Filippov DV, Zaja R, Feijs KL
  TITLE     Protein and RNA ADP-ribosylation detection is influenced by sample 
            preparation and reagents used.
  JOURNAL   Life Sci Alliance. 2022 Nov 11;6(1):e202201455. doi: 
            10.26508/lsa.202201455.
  PUBMED    36368907
REFERENCE   2  (bases 1 to 5890)
  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: "Life Sci 
            Alliance."; date: "2022-11-11"; volume: "6(1):e202201455. doi"; 
            pages: " 10.26508/lsa.202201455"
COMMENT     Sequence Label: pDEST17-DXO
FEATURES             Location/Qualifiers
     source          1..5890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..42
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        23..41
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             73..95
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             115..132
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    142..166
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1361..1385)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1438..1456)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1452..1499
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1865..1870,1871..1887,1888..1893))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1920..1938)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2006..2110
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2111..2179,2180..2971)
                     /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(2329..2348)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3142..3730
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3631..3650
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3884..3901
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3916..4055
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3981..4000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4141..4163
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4157..4348)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5810..5829
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cgagatctcg atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc
       61 tctagaaata attttgttta actttaagaa ggagatatac atatgtcgta ctaccatcac
      121 catcaccatc acctcgaatc aacaagtttg tacaaaaaag caggcttcat ggatcccagg
      181 gggaccaaga gaggagctga gaagacagag gtagctgagc ctcggaacaa actacctcgt
      241 ccagcacctt ctctgcccac agaccctgcc ctctactctg ggccctttcc tttctaccgg
      301 cgcccttcgg aactgggctg cttctccctg gatgctcaac gccagtacca tggagatgcc
      361 cgagccctgc gctactatag cccacccccc actaacggtc caggccccaa ctttgacctc
      421 agagacggat acccggatcg ataccagccc cgggacgagg aggtccagga aaggctggac
      481 cacctgctgt gctggctcct ggaacaccga ggccggttgg aggggggtcc aggctggctg
      541 gcagaggcca tagtgacgtg gcgggggcac ctgacaaaac tgctgacgac accgtatgag
      601 cggcaggagg gctggcagct ggcagcctcc cggttccagg gaacactata cctgagtgaa
      661 gtggagacac cgaacgctcg ggcccagagg cttgctcggc caccgctcct ccgggagctt
      721 atgtacatgg gatacaaatt tgagcagtac atgtgtgcag acaaacctgg aagctcccca
      781 gacccctctg gggaggttaa caccaacgtg gccttctgct ctgtgctacg cagccgcctg
      841 ggaagccacc ctctgctctt ctcaggggag gtagactgca cagaccccca agccccatcc
      901 acacagcccc caacctgcta tgtggagctc aagacctcca aggagatgca cagccctggc
      961 caatggagga gtttctacag acacaagctc ctgaaatggt gggctcagtc attcctccca
     1021 ggggtcccga atgttgttgc tggcttccgt aacccagacg gttttgtctc ttccctcaag
     1081 acctttccta ccatgaagat gtttgaatat gtcaggaatg accgtgacgg ctggaatccc
     1141 tctgtgtgca tgaacttctg tgccgccttc cttagctttg cccagagcac ggttgtccag
     1201 gatgacccca ggctcgttca tctcttctct tgggagcctg gcggcccagt caccgtgtct
     1261 gtacaccaag atgcacctta cgccttcctg cccatatggt atgtggaagc tatgactcag
     1321 gacctcccat caccccccaa gactccctct cccaaatagg acccagcttt cttgtacaaa
     1381 gtggttgatt cgaggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     1441 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     1501 tgaaaggagg aactatatcc ggatatccac aggacgggtg tggtcgccat gatcgcgtag
     1561 tcgatagtgg ctccaagtag cgaagcgagc aggactgggc ggcggccaaa gcggtcggac
     1621 agtgctccga gaacgggtgc gcatagaaat tgcatcaacg catatagcgc tagcagcacg
     1681 ccatagtgac tggcgatgct gtcggaatgg acgatatccc gcaagaggcc cggcagtacc
     1741 ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc
     1801 gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta
     1861 ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa gacgaaaggg
     1921 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     1981 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2041 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2101 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2161 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2221 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2281 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2341 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2401 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2461 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2521 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2581 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2641 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2701 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2761 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2821 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2881 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2941 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3001 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3061 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3121 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3181 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3241 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3301 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3361 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3421 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3481 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3541 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3601 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3661 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3721 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3781 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3841 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3901 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3961 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     4021 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     4081 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4141 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     4201 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     4261 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     4321 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     4381 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     4441 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     4501 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     4561 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     4621 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     4681 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     4741 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     4801 tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg
     4861 agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc
     4921 gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc
     4981 ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac
     5041 aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc
     5101 ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag
     5161 cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa
     5221 cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc
     5281 tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat
     5341 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     5401 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     5461 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     5521 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     5581 tgggttgaag gctctcaagg gcatcggtcg atcgacgctc tcccttatgc gactcctgca
     5641 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     5701 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     5761 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     5821 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     5881 cgtagaggat
//
