LOCUS       sequence-334844-        5440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged cDAXX .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5440)
  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 5440)
  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
FEATURES             Location/Qualifiers
     source          1..5440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(353..377)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(387..404)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             424..446
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(477..496)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(478..496)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(580..599)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             2061..2252
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2246..2268)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2354..2493
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2409..2428
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2508..2525)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2679..3267)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2759..2778)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3438..4229,4230..4298))
                     /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     4061..4080
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4299..4403)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4471..4489
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4516..4521,4522..4538,4539..4544)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(4910..4957)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4953..4971
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    5024..5048
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gtgctggaca ccatgccagc gccgttttcc agcacggtgg tgaagggttc ctcgctctgc
       61 ttccggctgc tgctgatgtc ggtttccacg ctgctgggtg tgtccagggg cagggcctcg
      121 atttccagct cgaacagctg ggacacgggg gattcctctt ccagggtcag ttcctcgggc
      181 tgttcgccat tagactcggc gtcgatggag ctgggggcca gaggttcttc gctcagcagg
      241 ctgggggaca cgattctgcc cttgttctgc tgctcgccgc tgcttctgga gatctgcttg
      301 ccgggttcca ggttcttctc gttgctgatc tgcaggctgc tcatggggct gaagcctgct
      361 tttttgtaca aacttgttga ttcgaggtga tggtgatggt gatggtagta cgacatatgt
      421 atatctcctt cttaaagtta aacaaaatta tttctagagg gaaaccgttg tggtctccct
      481 atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc ggacgcatcg
      541 tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg
      601 atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg
      661 tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg
      721 cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc
      781 ataagggaga gcgtcgatcg accgatgccc ttgagagcct tcaacccagt cagctccttc
      841 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
      901 ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg
      961 agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa
     1021 gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc
     1081 atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc
     1141 ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg
     1201 ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt
     1261 accagcctaa cttcgatcat tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg
     1321 agcacatgga acgggttggc atggattgta ggcgccgccc tataccttgt ctgcctcccc
     1381 gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc
     1441 tcgctaacgg attcaccact ccaagaattg gagccaatca attcttgcgg agaactgtga
     1501 atgcgcaaac caacccttgg cagaacatat ccatcgcgtc cgccatctcc agcagccgca
     1561 cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt
     1621 cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga
     1681 tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat
     1741 gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca
     1801 ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat
     1861 ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc
     1921 ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa
     1981 cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc
     2041 ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct
     2101 ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac
     2161 aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg
     2221 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     2281 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     2341 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     2401 taactatgcg gcatcagagc agattgtact gagagtgcac cattgcggtg tgaaataccg
     2461 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     2521 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2581 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2641 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2701 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2761 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2821 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2881 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2941 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3001 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3061 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     3121 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3181 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3241 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3301 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3361 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3421 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3481 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3541 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3601 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3661 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3721 gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg
     3781 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3841 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3901 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3961 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4021 atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc
     4081 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4141 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4201 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4261 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4321 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4381 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     4441 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt
     4501 caagaattct catgtttgac agcttatcat cgataagctt taatgcggta gtttatcaca
     4561 gttaaattgc taacgcagtc aggcaccgtg tatgaaatct aacaatgcgc tcatcgtcat
     4621 cctcggcacc gtcaccctgg atgctgtagg cataggcttg gttatgccgg tactgccggg
     4681 cctcttgcgg gatatcgtcc attccgacag catcgccagt cactatggcg tgctgctagc
     4741 gctatatgcg ttgatgcaat ttctatgcgc acccgttctc ggagcactgt ccgaccgctt
     4801 tggccgccgc ccagtcctgc tcgcttcgct acttggagcc actatcgact acgcgatcat
     4861 ggcgaccaca cccgtcctgt ggatatccgg atatagttcc tcctttcagc aaaaaacccc
     4921 tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg gtggcagcag
     4981 ccaactcagc ttcctttcgg gctttgttag cagcctcgaa tcaaccactt tgtacaagaa
     5041 agctgggtct tagtcgctgt cggacagcac gatgatctct tcggggtcgc actgggtggc
     5101 cacgcttgtc ttgctggtgc caggtctggg aggctggcta tgaggggtct ggctcagtct
     5161 ggcggggcta ggaatgctca ggctgctggt cacgaggccg tgagaagggc tatccactct
     5221 ggtgctgctg tcggccacag gagccagaga agccagaggg cttggggggc tgggcagggt
     5281 gctgatcttc ttgccgtttt tctcgtgcac gcttctctgc cgttccacgt agctgttgcc
     5341 cagagggccg ctgccggtct gcttcttctc tttccgggac ttcttgctgg gagggccaga
     5401 atcgccccag ttgtgagggg acacgccgcc attgaagctg
//
