LOCUS       sequence-334856-        5464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged cDAXX-0SB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5464)
  AUTHORS   Bouchard JJ, Otero JH, Scott DC, Szulc E, Martin EW, Sabri N, 
            Granata D, Marzahn MR, Lindorff-Larsen K, Salvatella X, Schulman BA,
            Mittag T
  TITLE     Cancer Mutations of the Tumor Suppressor SPOP Disrupt the Formation 
            of Active, Phase-Separated Compartments.
  JOURNAL   Mol Cell. 2018 Oct 4;72(1):19-36.e8. doi: 
            10.1016/j.molcel.2018.08.027. Epub 2018 Sep 20.
  PUBMED    30244836
REFERENCE   2  (bases 1 to 5464)
  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"; doi: 
            "10.1016/j.molcel.2018.08.027"; journalName: "Mol Cell"; date: 
            "2018-10-4- 4"; volume: "72"; issue: "1"; pages: "19-36.e8"
COMMENT     Sequence Label: pDEST17-cDAXX-0SB
FEATURES             Location/Qualifiers
     source          1..5464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(460..484)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(537..555)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      551..598
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(964..969,970..986,987..992))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1019..1037)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1105..1209
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1210..1278,1279..2070)
                     /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(1428..1447)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2241..2829
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2730..2749
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2983..3000
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3015..3154
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3080..3099)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3240..3262
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3256..3447)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4909..4928
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5012..5031
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5012..5030
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             5062..5084
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5104..5121
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    5131..5155
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5158..5178
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 ccatcaagca gtccgaggaa cccttcacca ccgtgctgga aaacggcgct ggcatggtca
       61 ccagcaccac cttcaatggc ggcgtgtccc cccacaactg gggcgattct ggccccccta
      121 gcaagaagtc ccggaaagag aagaagcaga ccggcagcgg ccctctgggc aacagctacg
      181 tggaacggca gagaagcgtg cacgagaaaa acggcaagaa gatcagcacc ctgcccagcc
      241 ccccaagccc tctggcttct ctggctcctg tgtcccccga tagcgccaga gtggatagcc
      301 ctagccacgg cctgagccca gtgtccctgt ctattcccag ccccgccaga ctgagccaga
      361 cccctcatag ccagcctccc agacctggca ccagcaagac aagcgtggcc acccagtgcg
      421 accccgaaga gatcatcgtg ctgtccgaca gcgactaaga cccagctttc ttgtacaaag
      481 tggttgattc gaggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
      541 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
      601 gaaaggagga actatatccg gatatccaca ggacgggtgt ggtcgccatg atcgcgtagt
      661 cgatagtggc tccaagtagc gaagcgagca ggactgggcg gcggccaaag cggtcggaca
      721 gtgctccgag aacgggtgcg catagaaatt gcatcaacgc atatagcgct agcagcacgc
      781 catagtgact ggcgatgctg tcggaatgga cgatatcccg caagaggccc ggcagtaccg
      841 gcataaccaa gcctatgcct acagcatcca gggtgacggt gccgaggatg acgatgagcg
      901 cattgttaga tttcatacac ggtgcctgac tgcgttagca atttaactgt gataaactac
      961 cgcattaaag cttatcgatg ataagctgtc aaacatgaga attcttgaag acgaaagggc
     1021 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     1081 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     1141 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     1201 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     1261 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     1321 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     1381 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     1441 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     1501 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     1561 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     1621 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     1681 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     1741 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     1801 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     1861 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     1921 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     1981 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     2041 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     2101 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     2161 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2221 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2281 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2341 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     2401 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2461 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2521 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2581 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2641 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2701 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2761 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2821 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2881 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     2941 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3001 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3061 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     3121 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     3181 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3241 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     3301 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     3361 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     3421 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     3481 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     3541 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     3601 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     3661 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     3721 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     3781 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     3841 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     3901 catgcgcacc cgtggccagg acccaacgct gcccgagatg cgccgcgtgc ggctgctgga
     3961 gatggcggac gcgatggata tgttctgcca agggttggtt tgcgcattca cagttctccg
     4021 caagaattga ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt gccgccggct
     4081 tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca
     4141 aggtataggg cggcgcctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca
     4201 taaatcgccg tgacgatcag cggtccaatg atcgaagtta ggctggtaag agccgcgagc
     4261 gatccttgaa gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac
     4321 gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct
     4381 cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg
     4441 gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg
     4501 tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg
     4561 tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag
     4621 aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact
     4681 gggttgaagg ctctcaaggg catcggtcga tcgacgctct cccttatgcg actcctgcat
     4741 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
     4801 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     4861 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     4921 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     4981 gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg gagaccacaa
     5041 cggtttccct ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgtcgtac
     5101 taccatcacc atcaccatca cctcgaatca acaagtttgt acaaaaaagc aggcttcgaa
     5161 aacctgtatt ttcagggcag cagccccatg agcagcctgc agatcagcaa cgagaagaac
     5221 ctggaacccg gcaagcagag ccccagatcc atcggcgagc agcagaacaa gggccggatc
     5281 gtgtctccca gcctgctgag cgaagaaccc ctggccccta gctccatcga cgccgagtct
     5341 aacggcgaac agcccgagga actgaccctg gaagaggaat cccccgtgtc ccagctgttc
     5401 gagctggaaa tcgaggccct gcccctggac acacccagca gcgtggaaac agacagcccc
     5461 cggt
//
