LOCUS       sequence-334845-        6619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression construct of GFP tagged cDAXX-0SB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6619)
  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 6619)
  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-0SB
FEATURES             Location/Qualifiers
     source          1..6619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(83..412)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      97..232
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(151..170)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     397..417
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(426..854)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(536..557)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     748..767
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    1056..1104
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1129..1148
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1188..1229)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    1237..1261
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2006..2030)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2046..2756,2757..2759,2760..2762))
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2088..2109)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2437..2456
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2696..2717
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2866..2890)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2887..3090)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2916..2936)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3091..3470
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3642..3661
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3736..3840
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3841..3909,3910..4701)
                     /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     complement(4059..4078)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4872..5460
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5361..5380
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5614..5631
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5748..5769
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5784..5789,5790..5807,5808..5814)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5822..5838
                     /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)."
     primer_bind     5827..5849
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5846..5862
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5846..5862
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5911..6032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5922..5941)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5976..5995
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6190..6588)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(join(6607..6619,1..35))
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
ORIGIN
        1 atgccgatat actatgccga tgattaattg tcaacacgtg ctgatcagat ccgaaaatgg
       61 atatacaagc tcccgggagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact
      121 acttctggaa tagctcagag gcagaggcgg cctcggcctc tgcataaata aaaaaaatta
      181 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
      241 ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
      301 ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca
      361 tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac acattccaca
      421 gaattaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
      481 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
      541 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
      601 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
      661 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
      721 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
      781 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
      841 gctacagggc gcgtggggat accccctaga gccccagctg cgcagatctg ctatggcagg
      901 gcctgccgcc ccgacgttgg ctgcgagccc tgggccttca cccgaacttg gggggtgggg
      961 tggggaaaag gaagaaacgc gggcgtattg gccccaatgg ggtctcggtg gggtatcgac
     1021 agagtgccag ccctgggacc gaaccccgcg tttatgaaca aacgacccaa cacccgtgcg
     1081 ttttattctg tctttttatt gccgtcatag cgcgggttcc ttccggtatt gtctccttcc
     1141 gtgtttcagt tagcctcccc cgtttaaact cattactaac cggtacgcgt agaatcgaga
     1201 ccgaggagag ggttagggat aggcttcccg ttatcaacca ctttgtacaa gaaagctggg
     1261 tcttagtcgc tgtcggacag cacgatgatc tcttcggggt cgcactgtgt ggccacgctg
     1321 gttctagagg tgccaggtct tggaggttgg ctgtgaggtg tctggctcag tctggctgga
     1381 gatgggatgc tcagagacac gggagacagt ccatggctag ggctatccac tctggcgcta
     1441 tcaggagaca ctggggccag agaagccaga ggacttggag gactaggcag tgtgctgatc
     1501 cgcctgccgt ttctctcgtg cacgcttctc tgtctttcca cgtagctgtt gcccagaggg
     1561 ccagatccgg tctgccttct ctctcttcta gaccgcctgc taggaggtcc agaatcgccc
     1621 cagttgtgag gggacactcc gccattaaag gtggtgctgg tgaccatgcc agcgccgttt
     1681 tccagcacgg tggtgaaagg ttcctcgctc tgtctgatgc tccgagggct gtctgtttcc
     1741 acgctgctgg gtgtatccag aggcagggcc tcgatttcca gctcgaacag ctgagacaca
     1801 ggggattcct cttccagggt cagttcctcg ggctgttcgc cattagactc ggcgtcgatg
     1861 gagctaggag ccagaggttc ttcgctcagc aggctgggag acacgattct gcctctgttc
     1921 tgctgctcgc cgatggatct agggctctgc ttgccaggtt ccaggttccg ctcgttgctg
     1981 atctgcaggc tgctcatggg gctgaagcct gcttttttgt acaaacttgt tgatgggctc
     2041 gagcccttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc
     2101 aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gggcggactg ggtgctcagg
     2161 tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg
     2221 tcggcgagct gcacgctgcc gtcctcgatg ttgtggcggg tcttgaagtt caccttgatg
     2281 ccgttcttct gcttgtcggc ggtgatatag accttgtggc tgttgtagtt gtactccagc
     2341 ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc
     2401 accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg
     2461 aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc
     2521 tgcttcatgt ggtcggggta gcgggcgaag cactgcacgc cgtaggtgaa ggtggtcacg
     2581 agggtgggcc agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg
     2641 ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg
     2701 ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc
     2761 atggtctcga ggatatcacc ggtagggatc ctctagagtc cggaggctgg atcggtcccg
     2821 gtgtcttcta tggaggtcaa aacagcgtgg atggcgtctc caggcgatct gacggttcac
     2881 taaacgagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     2941 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     3001 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     3061 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     3121 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     3181 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     3241 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     3301 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     3361 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     3421 taatgacccc gtaattgatt actattaata actagtcaat aatcaatgtc aacgcgtata
     3481 tctggcccgt acatcgcgaa gcagcgcaaa acgcctaacc ctaagcagat tcttcatgca
     3541 attgtcggtc aagccttgcc ttgttgtagc ttaaattttg ctcgcgcact actcagcgac
     3601 ctccaacaca caagcaggga gcagatactg gcttaactat gcggcatcag agcagattgt
     3661 actgagagtg caccataggg gatcgggaga tctcccgatc cgtcgacgtc aggtggcact
     3721 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3781 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3841 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3901 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3961 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4021 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4081 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4141 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4201 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4261 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4321 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4381 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4441 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4501 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4561 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4621 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4681 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4741 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4801 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4861 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4921 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4981 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5041 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5101 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5161 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5221 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5281 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5341 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5401 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5461 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5521 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5581 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     5641 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     5701 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     5761 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     5821 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
     5881 tagctagagg tcgacggtat acagacatga taagatacat tgatgagttt ggacaaacca
     5941 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     6001 ttgtaaccat tataagctgc aataaacaag ttggggtggg cgaagaactc cagcatgaga
     6061 tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt
     6121 tcatagaagg cggcggtgga atcgaaatct cgtagcacgt gtcagtcctg ctcctcggcc
     6181 acgaagtgct tagccctccc acacataacc agagggcagc aattcacgaa tcccaactgc
     6241 cgtcggctgt ccatcactgt ccttcactat ggctttgatc ccaggatgca gatcgagaag
     6301 cacctgtcgg caccgtccgc aggggctcaa gatgcccctg ttctcatttc cgatcgcgac
     6361 gatacaagtc aggttgccag ctgccgcagc agcagcagtg cccagcacca cgagttctgc
     6421 acaaggtccc ccagtaaaat gatatacatt gacaccagtg aagatgcggc cgtcgctaga
     6481 gagagctgcg ctggcgacgc tgtagtcttc agagatgggg atgctgttga ttgtagccgt
     6541 tgctctttca atgagggtgg attcttcttg agacaaaggc ttggccatgg tttagttcct
     6601 caccttgtcg tattatact
//
