LOCUS       sequence-334788-        6643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression construct of GFP tagged cDAXX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6643)
  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 6643)
  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: pcDNA6.2EmGFP-cDAXX
FEATURES             Location/Qualifiers
     source          1..6643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..172
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    230..254
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             257..277
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(1023..1047)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1055..1096
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1136..1155)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1180..1228
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1430..1858
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1517..1536)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1727..1748
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1867..1887)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1872..2201
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2052..2187
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2114..2133
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2249..2296
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2315..2713
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    2871..2992
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2908..2927)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2962..2981
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3041..3057)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3041..3057)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3054..3076)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3065..3081
                     /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(3089..3095,3096..3113,3114..3119))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3134..3155
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3272..3289)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3443..4031)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3523..3542)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4202..4993,4994..5062))
                     /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     4825..4844
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5063..5167)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5242..5261)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5433..5812
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5813..6016
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5967..5987
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6013..6037
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(6141..6143,6144..6146,6147..6643,1..214)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6186..6207)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6447..6466)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
       61 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
      121 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
      181 cgggatcact ctcggcatgg acgagctgta caagggctcg agcccatcaa caagtttgta
      241 caaaaaagca ggcttcgaaa acctgtattt tcagggcgcc agccccatgt cctcactaca
      301 gatctccaat gaaaagaacc tggaacctgg caaacagatc agcagatctt caggggagca
      361 gcaaaacaaa ggacgcatag tgtcaccatc gttactgtca gaagaacccc tggccccctc
      421 cagcatagat gctgaaagca atggagaaca gcctgaggag ctgaccctgg aggaagaaag
      481 ccctgtgtct cagctctttg agctagagat tgaagctttg cccctggata ccccttcctc
      541 tgtggagacg gacatctcct cttccaggaa gcaatcagag gagcccttca ccactgtctt
      601 agagaatgga gcaggcatgg tctcttctac ttccttcaat ggaggcgtct ctcctcacaa
      661 ctggggagat tctggtcccc cctgcaaaaa atctcggaag gagaagaagc aaacaggatc
      721 agggccatta ggaaacagct atgtggaaag gcaaaggtca gtgcatgaga agaatgggaa
      781 aaagatatgt accctgccca gcccaccttc ccccttggct tccttggccc cagttgctga
      841 ttcctccacg agggtggact ctcccagcca tggcctggtg accagctccc tctgcatccc
      901 ttctccagcc cggctgtccc aaacccccca ttcacagcct cctcggcctg gtacttgcaa
      961 gacaagtgtg gccacacaat gcgatccaga agagatcatc gtgctctcag actctgatta
     1021 ggacccagct ttcttgtaca aagtggttga taacgggaag cctatcccta accctctcct
     1081 cggtctcgat tctacgcgta ccggttagta atgagtttaa acgggggagg ctaactgaaa
     1141 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     1201 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     1261 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     1321 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     1381 ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt
     1441 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     1501 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     1561 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     1621 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     1681 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     1741 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     1801 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     1861 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     1921 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     1981 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2041 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2101 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     2161 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     2221 tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg
     2281 catagtataa tacgacaagg tgaggaacta aaccatggcc aagcctttgt ctcaagaaga
     2341 atccaccctc attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta
     2401 cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata
     2461 tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc
     2521 agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc
     2581 ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa
     2641 ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt
     2701 gtgggagggc taagcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
     2761 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     2821 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     2881 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2941 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     3001 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3061 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3121 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3181 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3241 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3301 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3361 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3421 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3481 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3541 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3601 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3661 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3721 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3781 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3841 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3901 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3961 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4021 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4081 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4141 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4201 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4261 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4321 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4381 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4441 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4501 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4561 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4621 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4681 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4741 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4801 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4861 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4921 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4981 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5041 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5101 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5161 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     5221 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     5281 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     5341 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     5401 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     5461 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     5521 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     5581 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     5641 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     5701 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     5761 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     5821 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     5881 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     5941 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     6001 ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg
     6061 ttttgacctc catagaagac accgggaccg atccagcctc cggactctag aggatcccta
     6121 ccggtgatat cctcgagacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     6181 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     6241 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     6301 tgcccgtgcc ctggcccacc ctcgtgacca ccttcaccta cggcgtgcag tgcttcgccc
     6361 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     6421 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     6481 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     6541 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaag gtctatatca
     6601 ccgccgacaa gcagaagaac ggcatcaagg tgaacttcaa gac
//
