LOCUS       sequence-334853-        6578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of mouse SPOP F133V.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6578)
  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 6578)
  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: SPOP-V5 (F133V)
FEATURES             Location/Qualifiers
     source          1..6578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             737..778
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1935..1952)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1941..2165
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2211..2639
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2298..2317)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2508..2529
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2648..2668)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2653..2982
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2833..2968
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2895..2914
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3049..3843
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3103..3122)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3713..3732
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4017..4138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4054..4073)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4108..4127
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4187..4203)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4187..4203)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4200..4222)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4211..4227
                     /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(4235..4241,4242..4259,4260..4265))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4280..4301
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4418..4435)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4589..5177)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4669..4688)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5348..6139,6140..6208))
                     /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     5971..5990
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6209..6313)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6388..6407)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
      721 ttctgcagat atcatgggta agcctatccc taaccctctc ctcggtctcg attctacgat
      781 gtcaagggtt ccaagtcctc cacctccggc agaaatgtcg agtggtcccg ttgccgagag
      841 ctggtgctat acacagatca aggtagtgaa gttctcctac atgtggacca tcaataactt
      901 tagtttttgc cgggaggaaa tgggtgaagt cataaaaagt tcaacttttt catcaggagc
      961 caatgataaa ctgaaatggt gtttgcgagt aaacccgaaa gggctagatg aagaaagcaa
     1021 agattacctg tcgctttacc tgttgttggt cagctgtcca aagagtgaag ttcgggcaaa
     1081 gttcaagttc tccatcctga atgccaaggg agaagaaacc aaagctatgg agagtcagcg
     1141 agcttatagg ttcgtgcaag gcaaagactg gggagtgaag aaattcatcc gcagagattt
     1201 tctcttggac gaggccaatg ggcttctccc tgatgacaag ctcaccctct tctgtgaggt
     1261 gagtgtagta caagactctg tcaatatttc tgggcagaat actatgaaca tggtaaaggt
     1321 tcctgaatgc cggttggcag atgagttagg agggctgtgg gagaactctc gattcaccga
     1381 ctgctgcttg tgtgtggctg gtcaagaatt ccaggctcac aaggctatct tagcagctcg
     1441 gtctccagtc ttcagtgcca tgtttgaaca tgaaatggag gagagcaaaa agaatcgggt
     1501 tgagattaac gacgtggagc ctgaggtgtt taaggagatg atgtgcttca tctacacggg
     1561 gaaggcgccg aacctcgaca agatggctga cgacttgctg gcagctgccg ataagtacgc
     1621 cctggagcgt ctgaaggtca tgtgtgagga cgccctctgc agtaacctct ctgtggagaa
     1681 cgctgcagag attctcatcc tggctgacct ccacagcgca gatcagctga agacgcaggc
     1741 agttgatttc atcaactatc atgcttcgga tgtcttggag acctctgggt ggaagtcaat
     1801 ggtggtgtca catccccact tagtggcaga agcataccgt tctctggctt cagcacagtg
     1861 ccctttcctg gggcccccac gcaaacgcct gaagcaatcc taatctagag ggcccgttta
     1921 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1981 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2041 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2101 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2161 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2221 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2281 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2341 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2401 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2461 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2521 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2581 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     2641 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2701 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2761 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2821 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2881 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     2941 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3001 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3061 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3121 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3181 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3241 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3301 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3361 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3421 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3481 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3541 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3601 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3661 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3721 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3781 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3841 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     3901 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     3961 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4021 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4081 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4141 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4201 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4261 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4321 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4381 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4441 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4501 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4561 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4621 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4681 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4741 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4801 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4861 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4921 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4981 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5041 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5101 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5161 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5221 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5281 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5341 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5401 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5461 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5521 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5581 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5641 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5701 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5761 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5821 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5881 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5941 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6001 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6061 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6121 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6181 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6241 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6301 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     6361 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6421 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     6481 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     6541 tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
